Compositions comprising bacterial strains

ABSTRACT

The invention provides compositions comprising bacterial strains for treating and preventing inflammatory and autoimmune diseases.

CROSS REFERENCE

This application is a continuation of U.S. application Ser. No.16/505,098, filed Jul. 8, 2019, which is a continuation of U.S.application Ser. No. 16/040,356, filed Jul. 19, 2018, now U.S. Pat. No.10,391,130, issued Aug. 27, 2019; which is a continuation of U.S.application Ser. No. 15/592,178, filed on May 10, 2017, now U.S. Pat.No. 10,058,574, issued Aug. 28, 2018, which is a continuation ofInternational Application No. PCT/GB2016/051768, filed Jun. 15, 2016;which claims the benefit of Great Britain Patent Application No.1510470.6, filed Jun. 15, 2015; Great Britain Patent Application No.1520510.7, filed Nov. 20, 2015; and Great Britain Patent Application No.1603786.3, filed Mar. 4, 2016, the entire contents of which are allincorporated by reference herein.

SEQUENCE LISTING

The instant application contains a Sequence Listing which has beensubmitted electronically in ASCII format and is hereby incorporated byreference in its entirety. Said ASCII copy, created on Mar. 31, 2020, isnamed 56708_713_304_SL.txt and is 5,218,304 bytes in size.

TECHNICAL FIELD

This invention is in the field of compositions comprising bacterialstrains isolated from the mammalian digestive tract and the use of suchcompositions in the treatment of disease.

BACKGROUND TO THE INVENTION

The human intestine is thought to be sterile in utero, but it is exposedto a large variety of maternal and environmental microbes immediatelyafter birth. Thereafter, a dynamic period of microbial colonization andsuccession occurs, which is influenced by factors such as delivery mode,environment, diet and host genotype, all of which impact upon thecomposition of the gut microbiota, particularly during early life.Subsequently, the microbiota stabilizes and becomes adult-like [1]. Thehuman gut microbiota contains more than 500-1000 different phylotypesbelonging essentially to two major bacterial divisions, theBacteroidetes and the Firmicutes [2]. The successful symbioticrelationships arising from bacterial colonization of the human gut haveyielded a wide variety of metabolic, structural, protective and otherbeneficial functions. The enhanced metabolic activities of the colonizedgut ensure that otherwise indigestible dietary components are degradedwith release of by-products providing an important nutrient source forthe host. Similarly, the immunological importance of the gut microbiotais well-recognized and is exemplified in germfree animals which have animpaired immune system that is functionally reconstituted following theintroduction of commensal bacteria [3-5].

Dramatic changes in microbiota composition have been documented ingastrointestinal disorders such as inflammatory bowel disease (IBD). Forexample, the levels of Clostridium cluster XIVa bacteria are reduced inIBD patients whilst numbers of E. coli are increased, suggesting a shiftin the balance of symbionts and pathobionts within the gut [6-9].Interestingly, this microbial dysbiosis is also associated withimbalances in T effector cell populations.

In recognition of the potential positive effect that certain bacterialstrains may have on the animal gut, various strains have been proposedfor use in the treatment of various diseases (see, for example,[10-13]). Also, certain strains, including mostly Lactobacillus andBifidobacterium strains, have been proposed for use in treating variousinflammatory and autoimmune diseases that are not directly linked to theintestines (see [14] and [15] for reviews). However, the relationshipbetween different diseases and different bacterial strains, and theprecise effects of particular bacterial strains on the gut and at asystemic level and on any particular types of diseases, are poorlycharacterised.

There is a requirement in the art for new methods of treatinginflammatory and autoimmune diseases. There is also a requirement forthe potential effects of gut bacteria to be characterised so that newtherapies using gut bacteria can be developed.

SUMMARY OF THE INVENTION

The inventors have developed new therapies for treating and preventinginflammatory and autoimmune diseases. In particular, the inventors havedeveloped new therapies for treating and preventing diseases andconditions mediated by IL-17 or the Th17 pathway. In particular, theinventors have identified that bacterial strains from the genusBacteroides can be effective for reducing the Th17 inflammatoryresponse. As described in the examples, oral administration ofcompositions comprising Bacteroides coprocola may reduce the severity ofthe inflammatory response, including the Th17 inflammatory response, inmouse models of asthma, rheumatoid arthritis and multiple sclerosis.

Therefore, in a first embodiment, the invention provides a compositioncomprising a bacterial strain of the genus Bacteroides, for use in amethod of treating or preventing a disease or condition mediated byIL-17 or the Th17 pathway. The inventors have identified that treatmentwith bacterial strains from this genus can reduce levels of cytokinesthat are part of the Th17 pathway, including IL-17, can alleviate theTh17 inflammatory response and can provide clinical benefits in mousemodels of inflammatory and autoimmune diseases mediated by IL-17 and theTh17 pathway.

In particular embodiments, the invention provides a compositioncomprising a bacterial strain of the genus Bacteroides, for use in amethod of treating or preventing a disease or condition selected fromthe group consisting of: multiple sclerosis; arthritis, such asrheumatoid arthritis, osteoarthritis, psoriatic arthritis, or juvenileidiopathic arthritis; neuromyelitis optica (Devic's disease); ankylosingspondylitis; spondyloarthritis; psoriasis; systemic lupus erythematosus;inflammatory bowel disease, such as Crohn's disease or ulcerativecolitis; celiac disease; asthma, such as allergic asthma or neutrophilicasthma; chronic obstructive pulmonary disease (COPD); cancer, such asbreast cancer, colon cancer, lung cancer or ovarian cancer; uveitis;scleritis; vasculitis; Behcet's disease; atherosclerosis; atopicdermatitis; emphysema; periodontitis; allergic rhinitis; and allograftrejection. The effect shown for the bacterial strains from the genusBacteroides on the Th17 inflammatory response may provide therapeuticbenefits for diseases and conditions mediated by IL-17 and the Th17pathway, such as those listed above.

In preferred embodiments, the invention provides a compositioncomprising a bacterial strain of the genus Bacteroides, for use in amethod of treating or preventing asthma, such as neutrophilic asthma orallergic asthma. The inventors have identified that treatment withBacteroides strains can reduce recruitment of neutrophils andeosinophils into the lungs, which can help treat or prevent asthma.Furthermore, the inventors have tested and demonstrated the efficacy ofBacteroides strains in mouse models of asthma. In certain embodiments,the composition is for use in a method of treating or preventingneutrophilic asthma or eosinophilic asthma. The effect shown for thecompositions of the invention on neutrophils and eosinophils mean thatthey may be particularly effective for treating or preventingneutrophilic asthma and eosinophilic asthma. Indeed, in certainembodiments, the composition is for use in a method of reducing aneutrophilic inflammatory response in the treatment or prevention ofasthma, or the composition is for use in a method of reducing aneosinophilic inflammatory response in the treatment or prevention ofasthma. In preferred embodiments, the invention provides a compositioncomprising a bacterial strain of the species Bacteroides coprocola, foruse in the treatment of asthma, and in particular neutrophilic asthma.Bacteroides coprocola is shown to have a particularly pronounced effecton neutrophils in asthma models and treatment with Bacteroides coprocolamay be particularly effective for treating neutrophilic asthma. Incertain embodiments, the invention provides a composition comprising abacterial strain of the species Bacteroides thetaiotaomicron for use inthe treatment of asthma, and in particular eosinophilic or allergicasthma. In certain embodiments, the invention provides a compositioncomprising a bacterial strain of the species Bacteroides fragilis foruse in the treatment of asthma, and in particular eosinophilic orallergic asthma.

In further preferred embodiments, the invention provides a compositioncomprising a bacterial strain of the genus Bacteroides, for use in amethod of treating or preventing rheumatoid arthritis. The inventorshave identified that treatment with Bacteroides strains can provideclinical benefits in a mouse model of rheumatoid arthritis and canreduce joint swelling. In preferred embodiments, the invention providesa composition comprising a bacterial strain of the species Bacteroidescoprocola, for use in the treatment of rheumatoid arthritis.Compositions using Bacteroides coprocola may be particularly effectivefor treating rheumatoid arthritis. In certain embodiments, the inventionprovides a composition comprising a bacterial strain of the speciesBacteroides thetaiotaomicron, for use in the treatment of rheumatoidarthritis. In certain embodiments, the invention provides a compositioncomprising a bacterial strain of the species Bacteroides fragilis, foruse in the treatment of rheumatoid arthritis.

In further preferred embodiments, the invention provides a compositioncomprising a bacterial strain of the genus Bacteroides, for use in amethod of treating or preventing multiple sclerosis. The inventors haveidentified that treatment with Bacteroides strains can reduce diseaseincidence and disease severity in a mouse model of multiple sclerosis.In preferred embodiments, the invention provides a compositioncomprising a bacterial strain of the species Bacteroides coprocola, foruse in the treatment of multiple sclerosis. Compositions usingBacteroides coprocola may be particularly effective for treatingmultiple sclerosis. In certain embodiments, the invention provides acomposition comprising a bacterial strain of the species Bacteroidesthetaiotaomicron, for use in the treatment of multiple sclerosis. Incertain embodiments, the invention provides a composition comprising abacterial strain of the species Bacteroides fragilis, for use in thetreatment of multiple sclerosis.

In further preferred embodiments, the invention provides a compositioncomprising a bacterial strain of the genus Bacteroides, for use in amethod of treating or preventing cancer, such as breast, lung or livercancer. Compositions comprising a bacterial strain of the genusBacteroides may reduce tumour growth in mouse models of breast, lung andliver cancer. In certain embodiments, the composition is for use in amethod of reducing tumour size or preventing tumour growth in thetreatment of cancer. In certain embodiments, the invention provides acomposition comprising a bacterial strain of the species Bacteroidescoprocola, for use in the treatment of cancer. In certain embodiments,the invention provides a composition comprising a bacterial strain ofthe species Bacteroides thetaiotaomicron, for use in the treatment ofcancer. In certain embodiments, the invention provides a compositioncomprising a bacterial strain of the species Bacteroides fragilis, foruse in the treatment of cancer.

In further preferred embodiments, the invention provides a compositioncomprising a bacterial strain of the genus Bacteroides, for use in amethod of treating or preventing uveitis, such as posterior uveitis. Incertain embodiments, the invention provides a composition comprising abacterial strain of the species Bacteroides coprocola, for use in thetreatment of uveitis. In certain embodiments, the invention provides acomposition comprising a bacterial strain of the species Bacteroidesthetaiotaomicron, for use in the treatment of uveitis. In certainembodiments, the invention provides a composition comprising a bacterialstrain of the species Bacteroides fragilis, for use in the treatment ofuveitis.

In certain embodiments, the compositions of the invention are for use ina method of reducing IL-17 production or reducing Th17 celldifferentiation in the treatment or prevention of a disease or conditionmediated by IL-17 or the Th17 pathway. In particular, the compositionsof the invention may be used in reducing IL-17 production or reducingTh17 cell differentiation in the treatment or prevention of asthma,rheumatoid arthritis or multiple sclerosis. Preferably, the inventionprovides compositions comprising a bacterial strain of the speciesBacteroides coprocola, for use in reducing IL-17 production or reducingTh17 cell differentiation in the treatment or prevention of asthma,rheumatoid arthritis or multiple sclerosis, or of asthma, rheumatoidarthritis, multiple sclerosis, uveitis or cancer. In certainembodiments, the invention provides compositions comprising a bacterialstrain of the species Bacteroides thetaiotaomicron, for use in reducingIL-17 production or reducing Th17 cell differentiation in the treatmentor prevention of asthma, rheumatoid arthritis, multiple sclerosis,uveitis or cancer. In certain embodiments, the invention providescompositions comprising a bacterial strain of the species Bacteroidesfragilis, for use in reducing IL-17 production or reducing Th17 celldifferentiation in the treatment or prevention of asthma, rheumatoidarthritis, multiple sclerosis, uveitis or cancer.

In certain embodiments, the composition is for use in a patient withelevated IL-17 levels or Th17 cells. The effect on the Th17 inflammatoryresponse shown for Bacteroides strains may be particularly beneficialfor such patients.

In preferred embodiments of the invention, the bacterial strain in thecomposition is of Bacteroides coprocola. Closely related strains mayalso be used, such as bacterial strains that have a 16s rRNA sequencethat is at least 95%, 96%, 97%, 98%, 99%, 99.5% or 99.9% identical tothe 16s rRNA sequence of a bacterial strain of Bacteroides coprocola.Preferably, the bacterial strain has a 16s rRNA sequence that is atleast 95%, 96%, 97%, 98%, 99%, 99.5% or 99.9% identical to SEQ ID NO:1,2, 3 or 4. Preferably, the sequence identity is to SEQ ID NO:4.Preferably, the bacterial strain for use in the invention has the 16srRNA sequence represented by SEQ ID NO:4.

In further preferred embodiments of the invention, the bacterial strainin the composition is of Bacteroides thetaiotaomicron. Closely relatedstrains may also be used, such as bacterial strains that have a 16s rRNAsequence that is at least 95%, 96%, 97%, 98%, 99%, 99.5% or 99.9%identical to the 16s rRNA sequence of a bacterial strain of Bacteroidesthetaiotaomicron. Preferably, the bacterial strain has a 16s rRNAsequence that is at least 95%, 96%, 97%, 98%, 99%, 99.5% or 99.9%identical to SEQ ID NO:5. Preferably, the bacterial strain for use inthe invention has the 16s rRNA sequence represented by SEQ ID NO:5.

In further preferred embodiments of the invention, the bacterial strainin the composition is of Bacteroides fragilis. Closely related strainsmay also be used, such as bacterial strains that have a genome withsequence identity to CR626927.1.

In certain embodiments, the composition of the invention is for oraladministration. Oral administration of the strains of the invention canbe effective for treating IL-17- or Th17 pathway-mediated diseases andconditions. Also, oral administration is convenient for patients andpractitioners and allows delivery to and/or partial or totalcolonisation of the intestine.

In certain embodiments, the composition of the invention comprises oneor more pharmaceutically acceptable excipients or carriers.

In certain embodiments, the composition of the invention comprises abacterial strain that has been lyophilised. Lyophilisation is aneffective and convenient technique for preparing stable compositionsthat allow delivery of bacteria.

In certain embodiments, the invention provides a food product comprisingthe composition as described above.

In certain embodiments, the invention provides a vaccine compositioncomprising the composition as described above.

Additionally, the invention provides a method of treating or preventinga disease or condition mediated by IL-17 or the Th17 pathway, comprisingadministering a composition comprising a bacterial strain of the genusBacteroides.

In developing the above invention, the inventors have identified andcharacterised a bacterial strain that is particularly useful fortherapy. The Bacteroides coprocola strain of the invention is shown tobe effective for treating the diseases described herein, such asarthritis, asthma and multiple sclerosis. Therefore, in another aspect,the invention provides a cell of the Bacteroides coprocola straindeposited under accession number NCIMB 42408, or a derivative thereof.The invention also provides compositions comprising such cells, orbiologically pure cultures of such cells. The invention also provides acell of the Bacteroides coprocola strain deposited under accessionnumber NCIMB 42408, or a derivative thereof, for use in therapy, inparticular for the diseases described herein.

BRIEF DESCRIPTION OF DRAWINGS

FIG. 1: Mouse model of house dust mite-induced asthma—Total BAL fluidcell counts.

FIG. 2: Mouse model of house dust mite-induced asthma—Total eosinophilcount in BALF.

FIG. 3: Mouse model of house dust mite-induced asthma—Proportion ofeosinophils in BALF.

FIG. 4: Mouse model of house dust mite-induced asthma—Total macrophagecount in BALF.

FIG. 5: Mouse model of house dust mite-induced asthma—Proportion ofmacrophages in BALF.

FIG. 6: Mouse model of house dust mite-induced asthma—Total neutrophilcount in BALF.

FIG. 7: Mouse model of house dust mite-induced asthma—Proportion ofneutrophils in BALF.

FIG. 8: Mouse model of house dust mite-induced asthma—Total lymphocytecount in BALF.

FIG. 9: Mouse model of house dust mite-induced asthma—Proportion oflymphocytes in BALF.

FIG. 10: Mouse model of severe neutrophilic asthma—Total BAL fluid cellcounts.

FIG. 11: Mouse model of severe neutrophilic asthma—Total eosinophilcount in BALF.

FIG. 12: Mouse model of severe neutrophilic asthma—Proportion ofeosinophils in BALF.

FIG. 13: Mouse model of severe neutrophilic asthma—Total macrophagecount in BALF.

FIG. 14: Mouse model of severe neutrophilic asthma—Proportion ofmacrophages in BALF.

FIG. 15: Mouse model of severe neutrophilic asthma—Total neutrophilcount in BALF.

FIG. 16: Mouse model of severe neutrophilic asthma—Proportion ofneutrophils in BALF.

FIG. 17: Mouse model of severe neutrophilic asthma—Total lymphocytecount in BALF.

FIG. 18: Mouse model of severe neutrophilic asthma—Proportion oflymphocytes in BALF.

FIG. 19: Mouse model of rheumatoid arthritis—Bodyweights, days −14 to 0.Data are presented as Mean±SEM percentages of the initial (Day −14)bodyweights. Statistical significance: ▴p<0.05 and ▴▴▴p<0.0001 whencompared to the vehicle-treated group.

FIG. 20: Mouse model of rheumatoid arthritis—Bodyweights, days 0 to 42.Data are presented as Mean±SEM percentages of the initial (Day 0)bodyweights. ▴p<0.05, ♦p<0.05, ▴▴▴p<0.001, ●●●●p<0.0001 when compared tothe vehicle-treated group.

FIG. 21: Mouse model of rheumatoid arthritis—Clinical Scores. Data arepresented as Mean±SEM. ****p<0.0001 when compared to Day 21 in thevehicle-treated group. ♦ p<0.05 when compared to the vehicle-treatedgroup on a given day.

FIG. 22: Mouse model of rheumatoid arthritis—Splenocyte proliferativeresponse to Collagen II. Media background subtracted[CII-stimulated—media background] counts per minute based on 3H-TdRincorporation. All data are presented as Mean±SEM.

FIG. 23: Mouse model of rheumatoid arthritis—Levels of IFNγ in tissueculture supernatants from Vehicle-treated group. Lines represent groupmedian values.

FIG. 24: Mouse model of rheumatoid arthritis—Levels of IL-17A in tissueculture supernatants from Vehicle-treated group. Lines represent groupmedian values.

FIG. 25: Mouse model of rheumatoid arthritis—Levels of IL-10 in tissueculture supernatants from Vehicle-treated group. Lines represent groupmedian values.

FIG. 26: Mouse model of rheumatoid arthritis—Levels of IL-6 in tissueculture supernatants from Vehicle-treated group. Lines represent groupmedian values.

FIG. 27: Mouse model of rheumatoid arthritis—Levels of cytokine intissue culture supernatants from biotherapeutic #675-treated group(Group 4). Lines represent group median values.

FIG. 28: Mouse model of house dust mite-induced asthma—Total IgE inSerum

FIG. 29: Mouse model of house dust mite-induced asthma—HDM specific IgG1in Serum

FIG. 30: Mouse model of house dust mite-induced asthma—Total IgE in BALF

FIG. 31: Mouse model of house dust mite-induced asthma—HDM specific IgG1in BALF

FIG. 32: Mouse model of house dust mite-induced asthma—HistologicalAnalysis—Mean Peribronchiolar Infiltration Score

FIG. 33: Mouse model of house dust mite-induced asthma—HistologicalAnalysis—Mean Perivascular Infiltration Score

FIG. 34: Mouse model of house dust mite-induced asthma—HistologicalAnalysis—Mean Inflammatory Score (Average of both Peribronchiolar andPerivascular Infiltration Score)

FIG. 35: Mouse model of house dust mite-induced asthma—HistologicalAnalysis—Mucus Score

FIG. 36: Mouse model of house dust mite-induced asthma—IL-9 level inlung tissue

FIG. 37: Mouse model of house dust mite-induced asthma—IL-1a level inlung tissue

FIG. 38: Mouse model of house dust mite-induced asthma—IFNγ level inlung tissue

FIG. 39: Mouse model of house dust mite-induced asthma—IL-17A level inlung tissue

FIG. 40: Mouse model of house dust mite-induced asthma—IL-4 level inlung tissue

FIG. 41: Mouse model of house dust mite-induced asthma—IL-5 level inlung tissue

FIG. 42: Mouse model of house dust mite-induced asthma—IL-1b level inlung tissue

FIG. 43: Mouse model of house dust mite-induced asthma—RANTES level inlung tissue

FIG. 44: Mouse model of house dust mite-induced asthma—MIP-1a level inlung tissue

FIG. 45: Mouse model of house dust mite-induced asthma—KC level in lungtissue

FIG. 46: Mouse model of house dust mite-induced asthma—MIP-2 level inlung tissue

FIG. 47: Mouse model of severe neutrophilic asthma—HDM specific IgG1 inSerum

FIG. 48: Mouse model of severe neutrophilic asthma—HDM specific IgG2a inSerum

FIG. 49: Mouse model of severe neutrophilic asthma—HDM specific IgG1 inBALF

FIG. 50: Mouse model of severe neutrophilic asthma—HDM specific IgG2a inBALF

FIG. 51: Mouse model of severe neutrophilic asthma—HistologicalAnalysis—Mean Peribronchiolar Infiltration Score

FIG. 52: Mouse model of severe neutrophilic asthma—HistologicalAnalysis—Mean Perivascular Infiltration Score

FIG. 53: Mouse model of severe neutrophilic asthma—HistologicalAnalysis—Mean Inflammatory Score (Average of both Peribronchiolar andPerivascular Infiltration Score)

FIG. 54: Mouse model of severe neutrophilic asthma—TNFα level in lungtissue

FIG. 55: Mouse model of severe neutrophilic asthma—IL-1a level in lungtissue

FIG. 56: Mouse model of severe neutrophilic asthma—IFNγ level in lungtissue

FIG. 57: Mouse model of severe neutrophilic asthma—IL-17F level in lungtissue

FIG. 58: Mouse model of severe neutrophilic asthma—IL-1b level in lungtissue

FIG. 59: Mouse model of severe neutrophilic asthma—RANTES level in lungtissue

FIG. 60: Mouse model of severe neutrophilic asthma—MIP-2 level in lungtissue

FIG. 61: Mouse model of severe neutrophilic asthma—KC level in lungtissue

FIG. 62: Mouse model of severe neutrophilic asthma—IL-17A level in lungtissue

FIG. 63: Mouse model of severe neutrophilic asthma—MIP-1a level in lungtissue

FIG. 64: Mouse model of severe neutrophilic asthma—IL-33 level in lungtissue

FIG. 65: Mouse model of rheumatoid arthritis—Visual Template forHistopathology Scoring. Representative images showing composite scoresfrom mouse tarsal joints in a collagen-induced arthritis study.

FIG. 66: Mouse model of rheumatoid arthritis—Histopathology:Inflammation Scores. Data are presented as Mean±SEM. **p<0.01 whencompared to the vehicle-treated group.

FIG. 67: Mouse model of rheumatoid arthritis—Histopathology:Vehicle-treated group Cartilage Scores. Data are presented as Mean±SEM.

FIG. 68: Mouse model of rheumatoid arthritis—Histopathology: BoneScores. Data are presented as Mean±SEM.

FIG. 69: Mouse model of rheumatoid arthritis—Histopathology: TotalScores. Data are presented as Mean±SEM.

FIG. 70: Mouse model of rheumatoid arthritis—Histopathology: Strain#675. Data are presented as Mean±SEM.

FIG. 71: Mouse model of rheumatoid arthritis—Histopathology:Representative Pictures. Animal ID (#n.n) and limb (R for right, L forleft) are indicated between brackets. Top left image (vehicle):extensive joint and bone destruction with inflammation and fibrosisextending to the peri-articular soft tissues. Lower image (strain #675):synovitis and bursitis extending focally to peri-articular tissues, mildarticular cartilage damage and intra-articular debris, bone structureunaffected.

FIG. 72: Mouse model of multiple sclerosis—clinical score.

FIG. 73: Mouse model of multiple sclerosis—disease incidence.

DISCLOSURE OF THE INVENTION

Bacterial Strains

The compositions of the invention comprise a bacterial strain of thegenus Bacteroides. The examples demonstrate that bacteria of this genusare useful for treating or preventing diseases and conditions mediatedby IL-17 or the Th17 pathway. The preferred bacterial strains are of thespecies Bacteroides coprocola. Further preferred bacterial strains areof the species Bacteroides thetaiotaomicron or Bacteroides fragilis.

Examples of Bacteroides species for use in the invention includeBacteroides massiliensis, Bacteroides coprocola, Bacteroidesthetaiotaomicron and Bacteroides caccae. A further example of aBacteroides species for use in the invention is Bacteroides fragilis.Bacteroides is a genus of Gram-negative, obligately anaerobic bacteria.Bacteroides species are non-endospore-forming bacilli, and may be eithermotile or non-motile, depending on the species. Bacteroides species makeup a substantial portion of the mammalian gastrointestinal flora and areessential for processing complex molecules.

Bacteroides coprocola cells cultivated on EG blood agar plates arestrictly anaerobic, non-spore-forming, non-motile and Gram-negative. Theshort rods or rod-shaped cells are about 0.8 μm in width and variable inlength, generally in the range 1-4 μm. Example strains of speciesBacteroides coprocola are described in [16]. The type strain, M16^(T)(=JCM 12979^(T)=DSM 17136^(T)), was isolated from faeces of a healthyhuman. Two additional strains [M11 (=JCM 12980) and M156 (=JCM 12981)]are included in this species. GenBank/EMBL/DDBJ accession numbers forthe 16S rRNA gene sequence of these Bacteroides coprocola strains areAB200223, AB200224 and AB200225 (disclosed herein as SEQ ID NO:1, SEQ IDNO:2 and SEQ ID NO:3).

The Bacteroides bacterium deposited under accession number NCIMB 42408was tested in the Examples and is also referred to herein as strain 675.A 16S rRNA sequence for the 675 strain that was tested is provided inSEQ ID NO:4. Strain 675 was deposited with the international depositaryauthority NCIMB, Ltd. (Ferguson Building, Aberdeen, AB21 9YA, Scotland)by 4D Pharma Research Ltd. (Life Sciences Innovation Building, Aberdeen,AB25 2ZS, Scotland) on 13 May 2015 as “Bacteroidales 675” and wasassigned accession number NCIMB 42408. The NCIMB deposit was made underthe terms of the Budapest Treaty. Maintenance of a viable culture isassured for 30 years from the date of deposit. All restrictions on theavailability to the public of the deposited microorganism will beirrevocably removed upon the granting of a patent for this application.

The genome of strain 675 comprises a chromosome and plasmid. Achromosome sequence for strain 675 is provided in SEQ ID NO:6. A plasmidsequence for strain 675 is provided in SEQ ID NO:7. These sequences weregenerated using the PacBio RS II platform.

Bacterial strains closely related to the strain tested in the examplesare also expected to be effective for treating or preventing diseasesand conditions mediated by IL-17 or the Th17 pathway. In certainembodiments, the bacterial strain for use in the invention has a 16srRNA sequence that is at least 95%, 96%, 97%, 98%, 99%, 99.5% or 99.9%identical to the 16s rRNA sequence of a bacterial strain of Bacteroidescoprocola. Preferably, the bacterial strain for use in the invention hasa 16s rRNA sequence that is at least 95%, 96%, 97%, 98%, 99%, 99.5% or99.9% identical to SEQ ID NO:1, 2, 3 or 4. Preferably, the sequenceidentity is to SEQ ID NO:4. Preferably, the bacterial strain for use inthe invention has the 16s rRNA sequence represented by SEQ ID NO:4.

A further preferred bacterial strain for use in the invention is theBacteroides thetaiotaomicron strain deposited under accession numberNCIMB 42341. This strain was deposited with the international depositaryauthority NCIMB, Ltd. (Ferguson Building, Aberdeen, AB21 9YA, Scotland)on 3 Dec. 2014.

Further preferred Bacteroides thetaiotaomicron strains for use in theinvention are the type strain ATCC 29148=CCUG 10774=CIP 104206=DSM2079=JCM 5827=NCTC 10582=VPI 5482 and strain WAL 2926=ATCC 29741. Afurther preferred Bacteroides thetaiotaomicron strain for use in theinvention is the strain described in EP1448995. The accession number forthe 16S rRNA gene sequence of Bacteroides thetaiotaomicron strain WAL2926 is M58763 (disclosed herein as SEQ ID NO:5).

In certain embodiments, the bacterial strain for use in the inventionhas a 16s rRNA sequence that is at least 95%, 96%, 97%, 98%, 99%, 99.5%or 99.9% identical to the 16s rRNA sequence of a bacterial strain ofBacteroides thetaiotaomicron. Preferably, the bacterial strain for usein the invention has a 16s rRNA sequence that is at least 95%, 96%, 97%,98%, 99%, 99.5% or 99.9% identical to SEQ ID NO:5. Preferably, thebacterial strain for use in the invention has the 16s rRNA sequencerepresented by SEQ ID NO:5.

A preferred Bacteroides fragilis strain for use in the invention is thetype strain ATCC 25285=CCUG 4856=CIP 77.16=DSM 2151=JCM 11019=LMG10263=NCTC 9343. The accession number for the Bacteroides fragilis NCTC9343 strain complete genome is CR626927.1 (version: CR626927.1GI:60491031).

In certain embodiments, the bacterial strain for use in the inventionhas a genome with sequence identity to CR626927.1. In preferredembodiments, the bacterial strain for use in the invention has a genomewith at least 90% sequence identity (e.g. at least 92%, 94%, 95%, 96%,97%, 98%, 99% or 100% sequence identity) to CR626927.1 across at least60% (e.g. at least 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, 99% or100%) of CR626927.1. For example, the bacterial strain for use in theinvention may have a genome with at least 90% sequence identity toCR626927.1 across 70% of CR626927.1, or at least 90% sequence identityto CR626927.1 across 80% of CR626927.1, or at least 90% sequenceidentity to CR626927.1 across 90% of CR626927.1, or at least 90%sequence identity to CR626927.1 across 100% of CR626927.1, or at least95% sequence identity to CR626927.1 across 70% of CR626927.1, or atleast 95% sequence identity to CR626927.1 across 80% of CR626927.1, orat least 95% sequence identity to CR626927.1 across 90% of CR626927.1,or at least 95% sequence identity to CR626927.1 across 100% ofCR626927.1, or at least 98% sequence identity to CR626927.1 across 70%of CR626927.1, or at least 98% sequence identity to CR626927.1 across80% of CR626927.1, or at least 98% sequence identity to CR626927.1across 90% of CR626927.1, or at least 98% sequence identity toCR626927.1 across 100% of CR626927.1.

In certain embodiments, the bacterial strain for use in the inventionhas a chromosome with sequence identity to SEQ ID NO:6. In preferredembodiments, the bacterial strain for use in the invention has achromosome with at least 90% sequence identity (e.g. at least 92%, 94%,95%, 96%, 97%, 98%, 99% or 100% sequence identity) to SEQ ID NO:6 acrossat least 60% (e.g. at least 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%,99% or 100%) of SEQ ID NO:6. For example, the bacterial strain for usein the invention may have a chromosome with at least 90% sequenceidentity to SEQ ID NO:6 across 70% of SEQ ID NO:6, or at least 90%sequence identity to SEQ ID NO:6 across 80% of SEQ ID NO:6, or at least90% sequence identity to SEQ ID NO:6 across 90% of SEQ ID NO:6, or atleast 90% sequence identity to SEQ ID NO:6 across 100% of SEQ ID NO:6,or at least 95% sequence identity to SEQ ID NO:6 across 70% of SEQ IDNO:6, or at least 95% sequence identity to SEQ ID NO:6 across 80% of SEQID NO:6, or at least 95% sequence identity to SEQ ID NO:6 across 90% ofSEQ ID NO:6, or at least 95% sequence identity to SEQ ID NO:6 across100% of SEQ ID NO:6, or at least 98% sequence identity to SEQ ID NO:6across 70% of SEQ ID NO:6, or at least 98% sequence identity to SEQ IDNO:6 across 80% of SEQ ID NO:6, or at least 98% sequence identity to SEQID NO:6 across 90% of SEQ ID NO:6, or at least 98% sequence identity toSEQ ID NO:6 across 100% of SEQ ID NO:6.

In certain embodiments, the bacterial strain for use in the inventionhas a plasmid with sequence identity to SEQ ID NO:7. In preferredembodiments, the bacterial strain for use in the invention has a plasmidwith at least 90% sequence identity (e.g. at least 92%, 94%, 95%, 96%,97%, 98%, 99% or 100% sequence identity) to SEQ ID NO:7 across at least60% (e.g. at least 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, 99% or100%) of SEQ ID NO:7. For example, the bacterial strain for use in theinvention may have a plasmid with at least 90% sequence identity to SEQID NO:7 across 70% of SEQ ID NO:7, or at least 90% sequence identity toSEQ ID NO:7 across 80% of SEQ ID NO:7, or at least 90% sequence identityto SEQ ID NO:7 across 90% of SEQ ID NO:7, or at least 90% sequenceidentity to SEQ ID NO:7 across 100% of SEQ ID NO:7, or at least 95%sequence identity to SEQ ID NO:7 across 70% of SEQ ID NO:7, or at least95% sequence identity to SEQ ID NO:7 across 80% of SEQ ID NO:7, or atleast 95% sequence identity to SEQ ID NO:7 across 90% of SEQ ID NO:7, orat least 95% sequence identity to SEQ ID NO:7 across 100% of SEQ IDNO:7, or at least 98% sequence identity to SEQ ID NO:7 across 70% of SEQID NO:7, or at least 98% sequence identity to SEQ ID NO:7 across 80% ofSEQ ID NO:7, or at least 98% sequence identity to SEQ ID NO:7 across 90%of SEQ ID NO:7, or at least 98% sequence identity to SEQ ID NO:7 across100% of SEQ ID NO:7.

In certain embodiments, the bacterial strain for use in the inventionhas a chromosome with sequence identity to SEQ ID NO:6 and a plasmidwith sequence identity to SEQ ID NO:7.

Bacterial strains that are biotypes of the bacterium deposited underaccession number 42408 are also expected to be effective for treating orpreventing diseases and conditions mediated by IL-17 or the Th17pathway. Bacterial strains that are biotypes of a bacterium depositedunder accession number NCIMB 42341, ATCC 29148 or ATCC 29741 are alsoexpected to be effective for treating or preventing diseases andconditions mediated by IL-17 or the Th17 pathway. A biotype is a closelyrelated strain that has the same or very similar physiological andbiochemical characteristics.

Strains that are biotypes of a bacterium deposited under accessionnumber NCIMB 42408, NCIMB 42341, ATCC 29148 or ATCC 29741 and that aresuitable for use in the invention may be identified by sequencing othernucleotide sequences for a bacterium deposited under accession numberNCIMB 42408, NCIMB 42341, ATCC 29148 or ATCC 29741. For example,substantially the whole genome may be sequenced and a biotype strain foruse in the invention may have at least 95%, 96%, 97%, 98%, 99%, 99.5% or99.9% sequence identity across at least 80% of its whole genome (e.g.across at least 85%, 90%, 95% or 99%, or across its whole genome). Othersuitable sequences for use in identifying biotype strains may includehsp60 or repetitive sequences such as BOX, ERIC, (GTG)₅, or REP or [17].Biotype strains may have sequences with at least 95%, 96%, 97%, 98%,99%, 99.5% or 99.9% sequence identity to the corresponding sequence of abacterium deposited under accession number NCIMB 42408, NCIMB 42341,ATCC 29148 or ATCC 29741.

Alternatively, strains that are biotypes of a bacterium deposited underaccession number NCIMB 42408, NCIMB 42341, ATCC 29148 or ATCC 29741 andthat are suitable for use in the invention may be identified by usingthe accession number NCIMB 42408 deposit or the accession number NCIMBdeposit 42341 or the accession number ATCC 29148 deposit or theaccession number ATCC 29741 deposit and restriction fragment analysisand/or PCR analysis, for example by using fluorescent amplified fragmentlength polymorphism (FAFLP) and repetitive DNA element (rep)-PCRfingerprinting, or protein profiling, or partial 16S or 23s rDNAsequencing. In preferred embodiments, such techniques may be used toidentify other Bacteroides coprocola or Bacteroides thetaiotaomicronstrains.

In certain embodiments, strains that are biotypes of a bacteriumdeposited under accession number NCIMB 42408, NCIMB 42341, ATCC 29148 orATCC 29741 and that are suitable for use in the invention are strainsthat provide the same pattern as a bacterium deposited under accessionnumber NCIMB 42408, NCIMB 42341, ATCC 29148 or ATCC 29741 when analysedby amplified ribosomal DNA restriction analysis (ARDRA), for examplewhen using Sau3AI restriction enzyme (for exemplary methods and guidancesee, for example,[18]). Alternatively, biotype strains are identified asstrains that have the same carbohydrate fermentation patterns as abacterium deposited under accession number NCIMB 42408, NCIMB 42341,ATCC 29148 or ATCC 29741.

Other Bacteroides strains that are useful in the compositions andmethods of the invention, such as biotypes of a bacterium depositedunder accession number NCIMB 42408, NCIMB 42341, ATCC 29148 or ATCC29741, may be identified using any appropriate method or strategy,including the assays described in the examples. For instance, strainsfor use in the invention may be identified by culturing in anaerobicYCFA and/or administering the bacteria to the type II collagen-inducedarthritis mouse model and then assessing cytokine levels. In particular,bacterial strains that have similar growth patterns, metabolic typeand/or surface antigens to a bacterium deposited under accession numberNCIMB 42408, NCIMB 42341, ATCC 29148 or ATCC 29741 may be useful in theinvention. A useful strain will have comparable immune modulatoryactivity to the NCIMB 42408, NCIMB 42341, ATCC 29148 or ATCC 29741strain. In particular, a biotype strain will elicit comparable effectson the asthma, arthritis and multiple sclerosis disease models andcomparable effects on cytokine levels to the effects shown in theExamples, which may be identified by using the culturing andadministration protocols described in the Examples.

A particularly preferred strain of the invention is the Bacteroidescoprocola strain deposited under accession number NCIMB 42408. This isthe exemplary 675 strain tested in the examples and shown to beeffective for treating disease. Therefore, the invention provides acell, such as an isolated cell, of the Bacteroides coprocola straindeposited under accession number NCIMB 42408, or a derivative thereof.The invention also provides a composition comprising a cell of theBacteroides coprocola strain deposited under accession number NCIMB42408, or a derivative thereof. The invention also provides abiologically pure culture of the Bacteroides coprocola strain depositedunder accession number NCIMB 42408. The invention also provides a cellof the Bacteroides coprocola strain deposited under accession numberNCIMB 42408, or a derivative thereof, for use in therapy, in particularfor the diseases described herein.

A derivative of the strain deposited under accession number NCIMB 42408,NCIMB 42341, ATCC 29148 or ATCC 29741 may be a daughter strain (progeny)or a strain cultured (subcloned) from the original. A derivative of astrain of the invention may be modified, for example at the geneticlevel, without ablating the biological activity. In particular, aderivative strain of the invention is therapeutically active. Aderivative strain will have comparable immune modulatory activity to theoriginal NCIMB 42408, NCIMB 42341, ATCC 29148 or ATCC 29741 strain. Inparticular, a derivative strain will elicit comparable effects on theasthma, arthritis and multiple sclerosis disease models and comparableeffects on cytokine levels to the effects shown in the Examples, whichmay be identified by using the culturing and administration protocolsdescribed in the Examples. A derivative of the NCIMB 42408 strain willgenerally be a biotype of the NCIMB 42408 strain. A derivative of theNCIMB 42341, ATCC 29148 or ATCC 29741 strain will generally be a biotypeof the NCIMB 42341, ATCC 29148 or ATCC 29741 strain.

References to cells of the Bacteroides coprocola strain deposited underaccession number NCIMB 42408 encompass any cells that have the samesafety and therapeutic efficacy characteristics as the strains depositedunder accession number NCIMB 42408, and such cells are encompassed bythe invention. References to cells of the Bacteroides thetaiotaomicronstrain deposited under accession numbers NCIMB 42341, ATCC 29148 or ATCC29741 encompass any cells that have the same safety and therapeuticefficacy characteristics as the strains deposited under accession numberNCIMB 42341, ATCC 29148 or ATCC 29741, and such cells are encompassed bythe invention.

In preferred embodiments, the bacterial strains in the compositions ofthe invention are viable and capable of partially or totally colonisingthe intestine.

Therapeutic Uses

As demonstrated in the examples, the bacterial compositions of theinvention are effective for reducing the Th17 inflammatory response. Inparticular, treatment with compositions of the invention achieves areduction in IL-17A levels and other Th17 pathway cytokines, andclinical improvements in animal models of conditions mediated by IL-17and the Th17 pathway. Therefore, the compositions of the invention maybe useful for treating or preventing inflammatory and autoimmunediseases, and in particular diseases or conditions mediated by IL-17. Inparticular, the compositions of the invention may be useful for reducingor preventing elevation of the IL-17 inflammatory response.

Th17 cells are a subset of T helper cells that produce, for example,IL-17A, IL17-F, IL-21 and IL-22. Th17 cell differentiation and IL-17expression may be driven by IL-23. These cytokines and others formimportant parts of the Th17 pathway, which is a well-establishedinflammatory signalling pathway that contributes to and underlies anumber of inflammatory and autoimmune diseases (as described in, forexample, [19-24]). Diseases wherein the Th17 pathway is activated areTh17 pathway-mediated diseases. Th17 pathway-mediated diseases can beameliorated or alleviated by repressing the Th17 pathway, which may bethrough a reduction in the differentiation of Th17 cells or a reductionin their activity or a reduction in the level of Th17 pathway cytokines.Diseases mediated by the Th17 pathway may be characterised by increasedlevels of cytokines produced by Th17 cells, such as IL-17A, IL-17F,IL-21, IL-22, IL-26, IL-9 (reviewed in [25]). Diseases mediated by theTh17 pathway may be characterised by increased expression ofTh-17-related genes, such as Stat3 or IL-23R. Diseases mediated by theTh17 pathway may be associated with increased levels of Th17 cells.

IL-17 is a pro-inflammatory cytokine that contributes to thepathogenesis of several inflammatory and autoimmune diseases andconditions. IL-17 as used herein may refer to any member of the IL-17family, including IL-17A, IL-17B, IL-17C, IL-17D, IL-17E, and IL-17F.IL-17-mediated diseases and conditions are characterised by highexpression of IL-17 and/or the accumulation or presence ofIL-17-positive cells in a tissue affected by the disease or condition.Similarly, IL-17-mediated diseases and conditions are diseases andconditions that are exacerbated by high IL-17 levels or an increase inIL-17 levels, and that are alleviated by low IL-17 levels or a reductionin IL-17 levels. The IL-17 inflammatory response may be local orsystemic.

Examples of diseases and conditions that may be mediated by IL-17 or theTh17 pathway include multiple sclerosis; arthritis, such as rheumatoidarthritis, osteoarthritis, psoriatic arthritis, or juvenile idiopathicarthritis; neuromyelitis optica (Devic's disease); ankylosingspondylitis; spondyloarthritis; psoriasis; systemic lupus erythematosus;inflammatory bowel disease, such as Crohn's disease or ulcerativecolitis; celiac disease; asthma, such as allergic asthma or neutrophilicasthma; chronic obstructive pulmonary disease (COPD); cancer, such asbreast cancer, colon cancer, lung cancer or ovarian cancer; uveitis;scleritis; vasculitis; Behcet's disease; atherosclerosis; atopicdermatitis; emphysema; periodontitis; allergic rhinitis; and allograftrejection. In preferred embodiments, the compositions of the inventionare used for treating or preventing one or more of these conditions ordiseases. In further preferred embodiments, these conditions or diseasesare mediated by IL-17 or the Th17 pathway.

In certain embodiments, the compositions of the invention are for use ina method of reducing IL-17 production or reducing Th17 celldifferentiation in the treatment or prevention of a disease or conditionmediated by IL-17 or the Th17 pathway. In certain embodiments, thecompositions of the invention are for use in treating or preventing aninflammatory or autoimmune disease, wherein said treatment or preventionis achieved by reducing or preventing elevation of the Th17 inflammatoryresponse. In certain embodiments, the compositions of the invention arefor use in treating a patient with an inflammatory or autoimmunedisease, wherein the patient has elevated IL-17 levels or elevated Th17cells or is exhibiting a Th17 inflammatory response. In certainembodiments, the patient may have been diagnosed with a chronicinflammatory or autoimmune disease or condition, or the composition ofthe invention may be for use in preventing an inflammatory or autoimmunedisease or condition developing into a chronic inflammatory orautoimmune disease or condition. In certain embodiments, the disease orcondition may not be responsive to treatment with TNF-α inhibitors.These uses of the invention may be applied to any of the specificdisease or conditions listed in the preceding paragraph.

IL-17 and the Th17 pathway are often associated with chronicinflammatory and autoimmune diseases, so the compositions of theinvention may be particularly useful for treating or preventing chronicdiseases or conditions as listed above. In certain embodiments, thecompositions are for use in patients with chronic disease. In certainembodiments, the compositions are for use in preventing the developmentof chronic disease.

The compositions of the invention may be useful for treating diseasesand conditions mediated by IL-17 or the Th17 pathway and for addressingthe Th17 inflammatory response, so the compositions of the invention maybe particularly useful for treating or preventing chronic disease,treating or preventing disease in patients that have not responded toother therapies (such as treatment with TNF-α inhibitors), and/ortreating or preventing the tissue damage and symptoms associated withIL-17 and Th17 cells. For example, IL-17 is known to activate matrixdestruction in cartilage and bone tissue and IL-17 has an inhibitoryeffect on matrix production in chondrocytes and osteoblasts, so thecompositions of the invention may be useful for treating or preventingbone erosion or cartilage damage.

In certain embodiments, treatment with compositions of the inventionprovides a reduction or prevents an elevation in IL-17 levels, inparticular IL-17A levels. In certain embodiments, treatment withcompositions of the invention provides a reduction or prevents anelevation in IFN-γ, IL-1β, RANTES, MIP-1α, IL-8 or IL-6 levels. Suchreduction or prevention of elevated levels of these cytokines may beuseful for treating or preventing inflammatory and autoimmune diseasesand conditions, in particular those mediated by IL-17 or the Th17pathway.

Asthma

In preferred embodiments, the compositions of the invention are for usein treating or preventing asthma. The examples demonstrate that thecompositions of the invention achieve a reduction in the recruitment ofneutrophils and/or eosinophils into the airways following sensitisationand challenge with house dust mite extract and so they may be useful inthe treatment or prevention of asthma. Asthma is a chronic diseasecharacterised by inflammation and restriction of the airways. Theinflammation in asthma may be mediated by IL-17 and/or Th17 cells, andso the compositions of the invention may be particularly effective forpreventing or treating asthma. The inflammation in asthma may bemediated by eosinophils and/or neutrophils.

In certain embodiments, the asthma is eosinophilic or allergic asthma.Eosinophilic and allergic asthma are characterised by increased numbersof eosinophils in peripheral blood and in airway secretions and isassociated pathologically with thickening of the basement membrane zoneand pharmacologically by corticosteroid responsiveness [26].Compositions that reduce or inhibit eosinophil recruitment or activationmay be useful for treating or preventing eosinophilic and allergicasthma.

In additional embodiments, the compositions of the invention are for usein treating or preventing neutrophilic asthma (or non-eosinophilicasthma). High neutrophil numbers are associated with severe asthma thatmay be insensitive to corticosteroid treatment. Compositions that reduceor inhibit neutrophil recruitment or activation may be useful fortreating or preventing neutrophilic asthma.

Eosinophilic and neutrophilic asthma are not mutually exclusiveconditions and treatments that help address either the eosinophil andneutrophil responses may be useful for treating asthma in general.

Increased IL-17 levels and activation of the Th17 pathway are associatedwith severe asthma, so the compositions of the invention may be usefulfor preventing the development of severe asthma or for treating severeasthma.

In certain embodiments, the compositions of the invention are for use inmethods reducing an eosinophilic inflammatory response in the treatmentor prevention of asthma, or for use in methods of reducing aneutrophilic inflammatory response in the treatment or prevention ofasthma. As noted above, high levels of eosinophils in asthma isassociated pathologically with thickening of the basement membrane zone,so reducing eosinophilic inflammatory response in the treatment orprevention of asthma may be able to specifically address this feature ofthe disease. Also, elevated neutrophils, either in combination withelevated eosinophils or in their absence, is associated with severeasthma and chronic airway narrowing. Therefore, reducing theneutrophilic inflammatory response may be particularly useful foraddressing severe asthma.

In certain embodiments, the compositions reduce peribronchiolarinfiltration in allergic asthma, or are for use in reducingperibronchiolar infiltration in the treatment of allergic asthma. Incertain embodiments, the compositions reduce peribronchiolar and/orperivascular infiltration in neutrophilic asthma, or are for use inreducing peribronchiolar and/or perivascular infiltration in thetreatment of allergic neutrophilic asthma.

In certain embodiments, treatment with compositions of the inventionprovides a reduction or prevents an elevation in IL-1β, IFNγ, RANTES,MIP-1α or IL-8 levels.

In certain embodiments, the compositions of the invention are for use ina method of treating asthma that results in a reduction of theeosinophilic and/or neutrophilic inflammatory response. In certainembodiments, the patient to be treated has, or has previously beenidentified as having, elevated neutrophil or eosinophil levels, forexample as identified through blood sampling or sputum analysis.

The compositions of the invention may be useful for preventing thedevelopment of asthma in a new-born when administered to the new-born,or to a pregnant woman. The compositions may be useful for preventingthe development of asthma in children. The compositions of the inventionmay be useful for treating or preventing adult-onset asthma. Thecompositions of the invention may be useful for managing or alleviatingasthma. The compositions of the invention may be particularly useful forreducing symptoms associated with asthma that is aggravated byallergens, such as house dust mites.

Treatment or prevention of asthma may refer to, for example, analleviation of the severity of symptoms or a reduction in the frequencyof exacerbations or the range of triggers that are a problem for thepatient.

Arthritis

In preferred embodiments, the compositions of the invention are for usein treating or preventing rheumatoid arthritis (RA). The examplesdemonstrate that the compositions of the invention achieve a reductionin the clinical signs of RA in a mouse model, reduce cartilage and bonedamage, and reduce the IL-17 inflammatory response, and so they may beuseful in the treatment or prevention of RA. RA is a systemicinflammatory disorder that primarily affects joints. RA is associatedwith an inflammatory response that results in swelling of joints,synovial hyperplasia, and destruction of cartilage and bone. IL-17 andTh17 cells may have a key role in RA, for example because IL-17 inhibitsmatrix production in chondrocytes and osteoblasts and activates theproduction and function of matrix metalloproteinases and because RAdisease activity is correlated to IL-17 levels and Th-17 cell numbers[27,28], so the compositions of the invention may be particularlyeffective for preventing or treating RA.

In certain embodiments, the compositions of the invention are for use inlowering IL-17 levels or preventing elevation of IL-17 levels in thetreatment or prevention of RA. In certain embodiments, treatment withcompositions of the invention provides a reduction or prevents anelevation in IL-17 levels, in particular IL-17A levels. In certainembodiments, treatment with compositions of the invention provides areduction or prevents an elevation in IFN-γ or IL-6 levels.

In certain embodiments, treatment with the compositions of the inventionresults in a reduction in the swelling of joints. In certainembodiments, the compositions of the invention are for use in patientswith swollen joints or patients identified as at risk of having swollenjoints. In certain embodiments, the compositions of the invention arefor use in a method of reducing joint swelling in RA.

In certain embodiments, treatment with the compositions of the inventionresults in a reduction in cartilage damage or bone damage. In certainembodiments, the compositions of the invention are for use in reducingor preventing cartilage or bone damage in the treatment of RA. Incertain embodiments, the compositions are for use in treating patientwith severe RA that are at risk of cartilage or bone damage.

Increased IL-17 levels and Th17 cell numbers are associated withcartilage and bone destruction in RA [27,28]. IL-17 is known to activatematrix destruction in cartilage and bone tissue and IL-17 has aninhibitory effect on matrix production in chondrocytes and osteoblasts.Therefore, in certain embodiments, the compositions of the invention arefor use in preventing bone erosion or cartilage damage in the treatmentof RA. In certain embodiments, the compositions are for use in treatingpatients that exhibit bone erosion or cartilage damage or patientsidentified as at risk of bone erosion or cartilage damage.

TNF-α is also associated with RA, but TNF-α is not involved in thepathogenesis of the later stages of the disease. In contrast, IL-17 hasa role throughout all stages of chronic disease [29]. Therefore, incertain embodiments the compositions of the invention are for use intreating chronic RA or late-stage RA, such as disease that includesjoint destruction and loss of cartilage. In certain embodiments, thecompositions of the invention are for treating patients that havepreviously received anti-TNF-α therapy. In certain embodiments, thepatients to be treated do not respond or no longer respond to anti-TNF-αtherapy.

The compositions of the invention may be useful for modulating apatient's immune system, so in certain embodiments the compositions ofthe invention are for use in preventing RA in a patient that has beenidentified as at risk of RA, or that has been diagnosed with early-stageRA. The compositions of the invention may be useful for preventing thedevelopment of RA.

The compositions of the invention may be useful for managing oralleviating RA. The compositions of the invention may be particularlyuseful for reducing symptoms associated with joint swelling or bonedestruction. Treatment or prevention of RA may refer to, for example, analleviation of the severity of symptoms or a reduction in the frequencyof exacerbations or the range of triggers that are a problem for thepatient.

Multiple Sclerosis

In preferred embodiments, the compositions of the invention are for usein treating or preventing multiple sclerosis. The examples demonstratethat the compositions of the invention achieve a reduction in thedisease incidence and disease severity in a mouse model of multiplesclerosis (the EAE model), and so they may be useful in the treatment orprevention of multiple sclerosis. Multiple sclerosis is an inflammatorydisorder associated with damage to the myelin sheaths of neurons,particularly in the brain and spinal column. Multiple sclerosis is achronic disease, which is progressively incapacitating and which evolvesin episodes. IL-17 and Th17 cells may have a key role in multiplesclerosis, for example because IL-17 levels may correlate with multiplesclerosis lesions, IL-17 can disrupt blood brain barrier endothelialcell tight junctions, and Th17 cells can migrate into the centralnervous system and cause neuronal loss [30,31]. Therefore, thecompositions of the invention may be particularly effective forpreventing or treating multiple sclerosis.

In certain embodiments, treatment with the compositions of the inventionresults in a reduction in disease incidence or disease severity. Incertain embodiments, the compositions of the invention are for use inreducing disease incidence or disease severity. In certain embodiments,treatment with the compositions of the invention prevents a decline inmotor function or results in improved motor function. In certainembodiments, the compositions of the invention are for use in preventinga decline in motor function or for use in improving motor function. Incertain embodiments, treatment with the compositions of the inventionprevents the development of paralysis. In certain embodiments, thecompositions of the invention are for use in preventing paralysis in thetreatment of multiple sclerosis.

The compositions of the invention may be useful for modulating apatient's immune system, so in certain embodiments the compositions ofthe invention are for use in preventing multiple sclerosis in a patientthat has been identified as at risk of multiple sclerosis, or that hasbeen diagnosed with early-stage multiple sclerosis or“relapsing-remitting” multiple sclerosis. The compositions of theinvention may be useful for preventing the development of sclerosis.Indeed, the examples show that administration of compositions of theinvention prevented the development of disease in many mice.

The compositions of the invention may be useful for managing oralleviating multiple sclerosis. The compositions of the invention may beparticularly useful for reducing symptoms associated with multiplesclerosis. Treatment or prevention of multiple sclerosis may refer to,for example, an alleviation of the severity of symptoms or a reductionin the frequency of exacerbations or the range of triggers that are aproblem for the patient.

Cancer

In preferred embodiments, the compositions of the invention are for usein treating or preventing cancer. IL-17 and the Th17 pathway havecentral roles in cancer development and progression, and so thecompositions of the invention may be useful for treating or preventingcancer.

Although the roles of IL-17 and Th17 cells in cancer are not fullyunderstood, numerous pro-tumour effects of IL-17 and Th17 cells areknown. For example, Th17 cells and IL-17 can promote angiogenesis,increase proliferation and survival of tumor cells and activatetumour-promoting transcription factors [32-34].

In certain embodiments, treatment with the compositions of the inventionresults in a reduction in tumour size or a reduction in tumour growth.In certain embodiments, the compositions of the invention are for use inreducing tumour size or reducing tumour growth. The compositions of theinvention may be effective for reducing tumour size or growth. Incertain embodiments, the compositions of the invention are for use inpatients with solid tumours. In certain embodiments, the compositions ofthe invention are for use in reducing or preventing angiogenesis in thetreatment of cancer. IL-17 and Th17 cells have central roles inangiogenesis. In certain embodiments, the compositions of the inventionare for use in preventing metastasis.

In certain embodiments, the compositions of the invention are for use intreating or preventing breast cancer. The compositions of the inventionmay be effective for treating breast cancer, and IL-17 and Th17 cellshave important roles in breast cancer [35]. In certain embodiments, thecompositions of the invention are for use in reducing tumour size,reducing tumour growth, or reducing angiogenesis in the treatment ofbreast cancer. In preferred embodiments the cancer is mammary carcinoma.In preferred embodiments the cancer is stage IV breast cancer.

In certain embodiments, the compositions of the invention are for use intreating or preventing lung cancer. The compositions of the inventionmay be effective for treating lung cancer, and IL-17 and Th17 cells haveimportant roles in lung cancer [36]. In certain embodiments, thecompositions of the invention are for use in reducing tumour size,reducing tumour growth, or reducing angiogenesis in the treatment oflung cancer. In preferred embodiments the cancer is lung carcinoma.

In certain embodiments, the compositions of the invention are for use intreating or preventing liver cancer. The compositions of the inventionmay be effective for treating liver cancer, and IL-17 and Th17 cellshave important roles in liver cancer [37]. In certain embodiments, thecompositions of the invention are for use in reducing tumour size,reducing tumour growth, or reducing angiogenesis in the treatment ofliver cancer. In preferred embodiments the cancer is hepatoma(hepatocellular carcinoma).

In certain embodiments, the compositions of the invention are for use intreating or preventing carcinoma. The compositions of the invention maybe particularly effective for treating carcinoma. In certainembodiments, the compositions of the invention are for use in treatingor preventing non-immunogenic cancer. The compositions of the inventionmay be effective for treating non-immunogenic cancers.

In further embodiments, the compositions of the invention are for use intreating or preventing acute lymphoblastic leukemia (ALL), acute myeloidleukemia, adrenocortical carcinoma, basal-cell carcinoma, bile ductcancer, bladder cancer, bone tumor, osteosarcoma/malignant fibroushistiocytoma, brainstem glioma, brain tumor, cerebellar astrocytoma,cerebral astrocytoma/malignant glioma, ependymoma, medulloblastoma,supratentorial primitive neuroectodermal tumors, breast cancer,bronchial adenomas/carcinoids, Burkitt's lymphoma, carcinoid tumor,cervical cancer, chronic lymphocytic leukemia, chronic myelogenousleukemia, chronic myeloproliferative disorders, colon cancer, cutaneousT-cell lymphoma, endometrial cancer, ependymoma, esophageal cancer,Ewing's sarcoma, intraocular melanoma, retinoblastoma, gallbladdercancer, gastric cancer, gastrointestinal carcinoid tumor,gastrointestinal stromal tumor (GIST), germ cell tumor, glioma,childhood visual pathway and hypothalamic, Hodgkin lymphoma, melanoma,islet cell carcinoma, Kaposi sarcoma, renal cell cancer, laryngealcancer, leukaemias, lymphomas, mesothelioma, neuroblastoma, non-Hodgkinlymphoma, oropharyngeal cancer, osteosarcoma, ovarian cancer, pancreaticcancer, parathyroid cancer, pharyngeal cancer, pituitary adenoma, plasmacell neoplasia, prostate cancer, renal cell carcinoma, retinoblastoma,sarcoma, testicular cancer, thyroid cancer, or uterine cancer.

The compositions of the invention may be particularly effective whenused in combination with further therapeutic agents. Theimmune-modulatory effects of the compositions of the invention may beeffective when combined with more direct anti-cancer agents. Therefore,in certain embodiments, the invention provides a composition comprisinga bacterial strain of the genus Bacteroides and an anticancer agent. Inpreferred embodiments the anticancer agent is an immune checkpointinhibitor, a targeted antibody immunotherapy, a CAR-T cell therapy, anoncolytic virus, or a cytostatic drug. In preferred embodiments, thecomposition comprises an anti-cancer agent selected from the groupconsisting of: Yervoy (ipilimumab, BMS); Keytruda (pembrolizumab,Merck); Opdivo (nivolumab, BMS); MEDI4736 (AZ/MedImmune); MPDL3280A(Roche/Genentech); Tremelimumab (AZ/MedImmune); CT-011 (pidilizumab,CureTech); BMS-986015 (lirilumab, BMS); MEDI0680 (AZ/MedImmune);MSB-0010718C (Merck); PF-05082566 (Pfizer); MEDI6469 (AZ/MedImmune);BMS-986016 (BMS); BMS-663513 (urelumab, BMS); IMP321 (Prima Biomed);LAG525 (Novartis); ARGX-110 (arGEN-X); PF-05082466 (Pfizer); CDX-1127(varlilumab; CellDex Therapeutics); TRX-518 (GITR Inc.); MK-4166(Merck); JTX-2011 (Jounce Therapeutics); ARGX-115 (arGEN-X); NLG-9189(indoximod, NewLink Genetics); INCB024360 (Incyte); IPH2201 (InnateImmotherapeutics/AZ); NLG-919 (NewLink Genetics); anti-VISTA (JnJ);Epacadostat (INCB24360, Incyte); F001287 (Flexus/BMS); CP 870893(University of Pennsylvania); MGA271 (Macrogenix); Emactuzumab(Roche/Genentech); Galunisertib (Eli Lilly); Ulocuplumab (BMS);BKT140/BL8040 (Biokine Therapeutics); Bavituximab (PeregrinePharmaceuticals); CC 90002 (Celgene); 852A (Pfizer); VTX-2337 (VentiRxPharmaceuticals); IMO-2055 (Hybridon, Idera Pharmaceuticals); LY2157299(Eli Lilly); EW-7197 (Ewha Women's University, Korea); Vemurafenib(Plexxikon); Dabrafenib (Genentech/GSK); BMS-777607 (BMS); BLZ945(Memorial Sloan-Kettering Cancer Centre); Unituxin (dinutuximab, UnitedTherapeutics Corporation); Blincyto (blinatumomab, Amgen); Cyramza(ramucirumab, Eli Lilly); Gazyva (obinutuzumab, Roche/Biogen); Kadcyla(ado-trastuzumab emtansine, Roche/Genentech); Perj eta (pertuzumab,Roche/Genentech); Adcetris (brentuximab vedotin, Takeda/Millennium);Arzerra (ofatumumab, GSK); Vectibix (panitumumab, Amgen); Avastin(bevacizumab, Roche/Genentech); Erbitux (cetuximab, BMS/Merck); Bexxar(tositumomab-I131, GSK); Zevalin (ibritumomab tiuxetan, Biogen); Campath(alemtuzumab, Bayer); Mylotarg (gemtuzumab ozogamicin, Pfizer);Herceptin (trastuzumab, Roche/Genentech); Rituxan (rituximab,Genentech/Biogen); volociximab (Abbvie); Enavatuzumab (Abbvie); ABT-414(Abbvie); Elotuzumab (Abbvie/BMS); ALX-0141 (Ablynx); Ozaralizumab(Ablynx); Actimab-C (Actinium); Actimab-P (Actinium); Milatuzumab-dox(Actinium); Emab-SN-38 (Actinium); Naptumonmab estafenatox (ActiveBiotech); AFM13 (Affimed); AFM11 (Affimed); AGS-16C3F (Agensys);AGS-16M8F (Agensys); AGS-22ME (Agensys); AGS-15ME (Agensys); GS-67E(Agensys); ALXN6000 (samalizumab, Alexion); ALT-836 (Altor Bioscience);ALT-801 (Altor Bioscience); ALT-803 (Altor Bioscience); AMG780 (Amgen);AMG 228 (Amgen); AMG820 (Amgen); AMG172 (Amgen); AMG595 (Amgen); AMG110(Amgen); AMG232 (adecatumumab, Amgen); AMG211 (Amgen/MedImmune);BAY20-10112 (Amgen/Bayer); Rilotumumab (Amgen); Denosumab (Amgen);AMP-514 (Amgen); MEDI575 (AZ/MedImmune); MEDI3617 (AZ/MedImmune);MEDI6383 (AZ/MedImmune); MEDI551 (AZ/MedImmune); Moxetumomab pasudotox(AZ/MedImmune); MEDI565 (AZ/MedImmune); MEDI0639 (AZ/MedImmune);MEDI0680 (AZ/MedImmune); MEDI562 (AZ/MedImmune); AV-380 (AVEO); AV203(AVEO); AV299 (AVEO); BAY79-4620 (Bayer); Anetumab ravtansine (Bayer);vantictumab (Bayer); BAY94-9343 (Bayer); Sibrotuzumab (BoehringerIngleheim); BI-836845 (Boehringer Ingleheim); B-701 (BioClin); BIIB015(Biogen); Obinutuzumab (Biogen/Genentech); BI-505 (Bioinvent); BI-1206(Bioinvent); TB-403 (Bioinvent); BT-062 (Biotest) BIL-010t (Biosceptre);MDX-1203 (BMS); MDX-1204 (BMS); Necitumumab (BMS); CAN-4 (Cantargia AB);CDX-011 (Celldex); CDX1401 (Celldex); CDX301 (Celldex); U3-1565 (DaiichiSankyo); patritumab (Daiichi Sankyo); tigatuzumab (Daiichi Sankyo);nimotuzumab (Daiichi Sankyo); DS-8895 (Daiichi Sankyo); DS-8873 (DaiichiSankyo); DS-5573 (Daiichi Sankyo); MORab-004 (Eisai); MORab-009 (Eisai);MORab-003 (Eisai); MORab-066 (Eisai); LY3012207 (Eli Lilly); LY2875358(Eli Lilly); LY2812176 (Eli Lilly); LY3012217(Eli Lilly); LY2495655 (EliLilly); LY3012212 (Eli Lilly); LY3012211 (Eli Lilly); LY3009806 (EliLilly); cixutumumab (Eli Lilly); Flanvotumab (Eli Lilly); IMC-TR1 (EliLilly); Ramucirumab (Eli Lilly); Tabalumab (Eli Lilly); Zanolimumab(Emergent Biosolution); FG-3019 (FibroGen); FPA008 (Five PrimeTherapeutics); FP-1039 (Five Prime Therapeutics); FPA144 (Five PrimeTherapeutics); catumaxomab (Fresenius Biotech); IMAB362 (Ganymed);IMAB027 (Ganymed); HuMax-CD74 (Genmab); HuMax-TFADC (Genmab); GS-5745(Gilead); GS-6624 (Gilead); OMP-21M18 (demcizumab, GSK); mapatumumab(GSK); IMGN289 (ImmunoGen); IMGN901 (ImmunoGen); IMGN853 (Immuno Gen);IMGN529 (Immuno Gen); IM MU-130 (Immunomedics); milatuzumab-dox(Immunomedics); IM MU-115 (Immunomedics); IMMU-132 (Immunomedics);IMMU-106 (Immunomedics); IMMU-102 (Immunomedics); Epratuzumab(Immunomedics); Clivatuzumab (Immunomedics); IPH41 (InnateImmunotherapeutics); Daratumumab (Janssen/Genmab); CNTO-95 (Intetumumab,Janssen); CNTO-328 (siltuximab, Janssen); KB004 (KaloBios);mogamulizumab (Kyowa Hakko Kirrin); KW-2871 (ecromeximab, Life Science);Sonepcizumab (Lpath); Margetuximab (Macrogenics); Enoblituzumab(Macrogenics); MGD006 (Macrogenics); MGF007 (Macrogenics); MK-0646(dalotuzumab, Merck); MK-3475 (Merck); Sym004 (Symphogen/Merck Serono);DI17E6 (Merck Serono); MOR208 (Morphosys); MOR202 (Morphosys); Xmab5574(Morphosys); BPC-1C (ensituximab, Precision Biologics); TAS266(Novartis); LFA102 (Novartis); BHQ880 (Novartis/Morphosys); QGE031(Novartis); HCD122 (lucatumumab, Novartis); LJM716 (Novartis); AT355(Novartis); OMP-21M18 (Demcizumab, OncoMed); OMP52M51 (Oncomed/GSK);OMP-59R5 (Oncomed/GSK); vantictumab (Oncomed/Bayer); CMC-544 (inotuzumabozogamicin, Pfizer); PF-03446962 (Pfizer); PF-04856884 (Pfizer);PSMA-ADC (Progenics); REGN1400 (Regeneron); REGN910 (nesvacumab,Regeneron/Sanofi); REGN421 (enoticumab, Regeneron/Sanofi); RG7221,RG7356, RG7155, RG7444, RG7116, RG7458, RG7598, RG7599, RG7600, RG7636,RG7450, RG7593, RG7596, DCDS3410A, RG7414 (parsatuzumab), RG7160(imgatuzumab), RG7159 (obintuzumab), RG7686, RG3638 (onartuzumab),RG7597 (Roche/Genentech); SAR307746 (Sanofi); SAR566658 (Sanofi);SAR650984 (Sanofi); SAR153192 (Sanofi); SAR3419 (Sanofi); SAR256212(Sanofi), SGN-LIV1A (lintuzumab, Seattle Genetics); SGN-CD33A (SeattleGenetics); SGN-75 (vorsetuzumab mafodotin, Seattle Genetics); SGN-19A(Seattle Genetics) SGN-CD70A (Seattle Genetics); SEA-CD40 (SeattleGenetics); ibritumomab tiuxetan (Spectrum); MLN0264 (Takeda); ganitumab(Takeda/Amgen); CEP-37250 (Teva); TB-403 (Thrombogenic); VB4-845(Viventia); Xmab2512 (Xencor); Xmab5574 (Xencor); nimotuzumab (YMBiosciences); Carlumab (Janssen); NY-ESO TCR (Adaptimmune); MAGE-A-10TCR (Adaptimmune); CTL019 (Novartis); JCAR015 (Juno Therapeutics);KTE-C19 CAR (Kite Pharma); UCART19 (Cellectis); BPX-401 (BellicumPharmaceuticals); BPX-601 (Bellicum Pharmaceuticals); ATTCK20 (UnumTherapeutics); CAR-NKG2D (Celyad); Onyx-015 (Onyx Pharmaceuticals); H101(Shanghai Sunwaybio); DNX-2401 (DNAtrix); VCN-01 (VCN Biosciences);Colo-Ad1 (PsiOxus Therapeutics); ProstAtak (Advantagene); Oncos-102(Oncos Therapeutics); CG0070 (Cold Genesys); Pexa-vac (JX-594, JennerexBiotherapeutics); GL-ONC1 (Genelux); T-VEC (Amgen); G207 (Medigene);HF10 (Takara Bio); SEPREHVIR (HSV1716, Virttu Biologics); OrienX010(OrienGene Biotechnology); Reolysin (Oncolytics Biotech); SVV-001(Neotropix); Cacatak (CVA21, Viralytics); Alimta (Eli Lilly), cisplatin,oxaliplatin, irinotecan, folinic acid, methotrexate, cyclophosphamide,5-fluorouracil, Zykadia (Novartis), Tafinlar (GSK), Xalkori (Pfizer),Iressa (AZ), Gilotrif (Boehringer Ingelheim), Tarceva (Astellas Pharma),Halaven (Eisai Pharma), Veliparib (Abbvie), AZD9291 (AZ), Alectinib(Chugai), LDK378 (Novartis), Genetespib (Synta Pharma),Tergenpumatucel-L (NewLink Genetics), GV1001 (Kael-GemVax), Tivantinib(ArQule); Cytoxan (BMS); Oncovin (Eli Lilly); Adriamycin (Pfizer);Gemzar (Eli Lilly); Xeloda (Roche); Ixempra (BMS); Abraxane (Celgene);Trelstar (Debiopharm); Taxotere (Sanofi); Nexavar (Bayer); IMMU-132(Immunomedics); E7449 (Eisai); Thermodox (Celsion); Cometriq (Exellxis);Lonsurf (Taiho Pharmaceuticals); Camptosar (Pfizer); UFT (TaihoPharmaceuticals); and TS-1 (Taiho Pharmaceuticals).

Uveitis

In further preferred embodiments, the compositions of the invention arefor use in treating or preventing uveitis. The compositions of theinvention may achieve a reduction in disease incidence and diseaseseverity in an animal model of uveitis and so they may be useful in thetreatment or prevention of uveitis. Uveitis is inflammation of the uveaand can result in retinal tissue destruction. It can present indifferent anatomical forms (anterior, intermediate, posterior ordiffuse) and result from different, but related, causes, includingsystemic autoimmune disorders. IL-17 and the Th17 pathway are centrallyinvolved in uveitis, so the compositions of the invention may beparticularly effective for preventing or treating uveitis. References[38-45] describe elevated serum levels of interleukin-17A in uveitispatients, specific association of IL17A genetic variants withpanuveitis, the role of Th17-associated cytokines in the pathogenesis ofexperimental autoimmune uveitis, the imbalance between Th17 Cells andregulatory T Cells during monophasic experimental autoimmune uveitis,the up-regulation of IL-17A in patients with uveitis and activeAdamantiades-Behcet and Vogt-Koyanagi-Harada (VKH) diseases, thetreatment of non-infectious uveitis with secukinumab (anti-IL-17Aantibody), and Th17 in uveitic eyes.

In certain embodiments, the uveitis is posterior uveitis. Posterioruveitis presents primarily with inflammation of the retina and choroidand the compositions of the invention may be effective for reducingretinal inflammation and damage.

In certain embodiments, treatment with the compositions of the inventionresults in a reduction in retinal damage. In certain embodiments, thecompositions of the invention are for use in reducing or preventingretinal damage in the treatment of uveitis. In certain embodiments, thecompositions are for use in treating patients with severe uveitis thatare at risk of retinal damage. In certain embodiments, treatment withthe compositions of the invention results in a reduction in optic discinflammation. In certain embodiments, the compositions of the inventionare for use in reducing or preventing optic disc inflammation. Incertain embodiments, treatment with the compositions of the inventionresults in a reduction in retinal tissue infiltration by inflammatorycells. In certain embodiments, the compositions of the invention are foruse in reducing retinal tissue infiltration by inflammatory cells. Incertain embodiments, treatment with the compositions of the inventionresults in vision being maintained or improved. In certain embodiments,the compositions of the invention are for use in maintaining orimproving vision.

In certain embodiments, the compositions are for use in treating orpreventing uveitis associated with a non-infectious or autoimmunedisease, such as Behcet disease, Crohn's disease, Fuchs heterochromiciridocyclitis, granulomatosis with polyangiitis, HLA-B27 relateduveitis, juvenile idiopathic arthritis, sarcoidosis, spondyloarthritis,sympathetic ophthalmia, tubulointerstitial nephritis and uveitissyndrome or Vogt-Koyanagi-Harada syndrome. IL-17A has been shown to beinvolved in, for example, Behcet and Vogt-Koyanagi-Harada diseases.

Treatment or prevention of uveitis may refer to, for example, analleviation of the severity of symptoms or a prevention of relapse.

Modes of Administration

Preferably, the compositions of the invention are to be administered tothe gastrointestinal tract in order to enable delivery to and/or partialor total colonisation of the intestine with the bacterial strain of theinvention. Generally, the compositions of the invention are administeredorally, but they may be administered rectally, intranasally, or viabuccal or sublingual routes.

In certain embodiments, the compositions of the invention may beadministered as a foam, as a spray or a gel.

In certain embodiments, the compositions of the invention may beadministered as a suppository, such as a rectal suppository, for examplein the form of a theobroma oil (cocoa butter), synthetic hard fat (e.g.suppocire, witepsol), glycero-gelatin, polyethylene glycol, or soapglycerin composition.

In certain embodiments, the composition of the invention is administeredto the gastrointestinal tract via a tube, such as a nasogastric tube,orogastric tube, gastric tube, jejunostomy tube (J tube), percutaneousendoscopic gastrostomy (PEG), or a port, such as a chest wall port thatprovides access to the stomach, jejunum and other suitable access ports.

The compositions of the invention may be administered once, or they maybe administered sequentially as part of a treatment regimen. In certainembodiments, the compositions of the invention are to be administereddaily.

In certain embodiments of the invention, treatment according to theinvention is accompanied by assessment of the patient's gut microbiota.Treatment may be repeated if delivery of and/or partial or totalcolonisation with the strain of the invention is not achieved such thatefficacy is not observed, or treatment may be ceased if delivery and/orpartial or total colonisation is successful and efficacy is observed.

In certain embodiments, the composition of the invention may beadministered to a pregnant animal, for example a mammal such as a humanin order to prevent an inflammatory or autoimmune disease developing inher child in utero and/or after it is born.

The compositions of the invention may be administered to a patient thathas been diagnosed with a disease or condition mediated by IL-17 or theTh17 pathway, or that has been identified as being at risk of a diseaseor condition mediated by IL-17 or the Th17 pathway. The compositions mayalso be administered as a prophylactic measure to prevent thedevelopment of diseases or conditions mediated by IL-17 or the Th17pathway in a healthy patient.

The compositions of the invention may be administered to a patient thathas been identified as having an abnormal gut microbiota. For example,the patient may have reduced or absent colonisation by Bacteroides, andin particular Bacteroides coprocola.

The compositions of the invention may be administered as a food product,such as a nutritional supplement.

Generally, the compositions of the invention are for the treatment ofhumans, although they may be used to treat animals including monogastricmammals such as poultry, pigs, cats, dogs, horses or rabbits. Thecompositions of the invention may be useful for enhancing the growth andperformance of animals. If administered to animals, oral gavage may beused.

Compositions

Generally, the composition of the invention comprises bacteria. Inpreferred embodiments of the invention, the composition is formulated infreeze-dried form. For example, the composition of the invention maycomprise granules or gelatin capsules, for example hard gelatincapsules, comprising a bacterial strain of the invention.

Preferably, the composition of the invention comprises lyophilisedbacteria. Lyophilisation of bacteria is a well-established procedure andrelevant guidance is available in, for example, references [46-48].

Alternatively, the composition of the invention may comprise a live,active bacterial culture.

In preferred embodiments, the composition of the invention isencapsulated to enable delivery of the bacterial strain to theintestine. Encapsulation protects the composition from degradation untildelivery at the target location through, for example, rupturing withchemical or physical stimuli such as pressure, enzymatic activity, orphysical disintegration, which may be triggered by changes in pH. Anyappropriate encapsulation method may be used. Exemplary encapsulationtechniques include entrapment within a porous matrix, attachment oradsorption on solid carrier surfaces, self-aggregation by flocculationor with cross-linking agents, and mechanical containment behind amicroporous membrane or a microcapsule. Guidance on encapsulation thatmay be useful for preparing compositions of the invention is availablein, for example, references [49] and [50].

The composition may be administered orally and may be in the form of atablet, capsule or powder. Encapsulated products are preferred becauseBacteroides are anaerobes. Other ingredients (such as vitamin C, forexample), may be included as oxygen scavengers and prebiotic substratesto improve the delivery and/or partial or total colonisation andsurvival in vivo. Alternatively, the probiotic composition of theinvention may be administered orally as a food or nutritional product,such as milk or whey based fermented dairy product, or as apharmaceutical product.

The composition may be formulated as a probiotic.

A composition of the invention includes a therapeutically effectiveamount of a bacterial strain of the invention. A therapeuticallyeffective amount of a bacterial strain is sufficient to exert abeneficial effect upon a patient. A therapeutically effective amount ofa bacterial strain may be sufficient to result in delivery to and/orpartial or total colonisation of the patient's intestine.

A suitable daily dose of the bacteria, for example for an adult human,may be from about 1×10³ to about 1×10¹¹ colony forming units (CFU); forexample, from about 1×10⁷ to about 1×10¹⁰ CFU; in another example fromabout 1×10⁶ to about 1×10¹⁰ CFU.

In certain embodiments, the composition contains the bacterial strain inan amount of from about 1×10⁶ to about 1×10¹¹ CFU/g, respect to theweight of the composition; for example, from about 1×10⁸ to about 1×10¹⁰CFU/g. The dose may be, for example, 1 g, 3 g, 5 g, and 10 g.

Typically, a probiotic, such as the composition of the invention, isoptionally combined with at least one suitable prebiotic compound. Aprebiotic compound is usually a non-digestible carbohydrate such as anoligo- or polysaccharide, or a sugar alcohol, which is not degraded orabsorbed in the upper digestive tract. Known prebiotics includecommercial products such as inulin and transgalacto-oligosaccharides.

In certain embodiments, the probiotic composition of the presentinvention includes a prebiotic compound in an amount of from about 1 toabout 30% by weight, respect to the total weight composition, (e.g. from5 to 20% by weight). Carbohydrates may be selected from the groupconsisting of: fructo-oligosaccharides (or FOS), short-chainfructo-oligosaccharides, inulin, isomalt-oligosaccharides, pectins,xylo-oligosaccharides (or XOS), chitosan-oligosaccharides (or COS),beta-glucans, arable gum modified and resistant starches, polydextrose,D-tagatose, acacia fibers, carob, oats, and citrus fibers. In oneaspect, the prebiotics are the short-chain fructo-oligosaccharides (forsimplicity shown herein below as FOSs-c.c); said FOSs-c.c. are notdigestible carbohydrates, generally obtained by the conversion of thebeet sugar and including a saccharose molecule to which three glucosemolecules are bonded.

The compositions of the invention may comprise pharmaceuticallyacceptable excipients or carriers. Examples of such suitable excipientsmay be found in the reference [51]. Acceptable carriers or diluents fortherapeutic use are well known in the pharmaceutical art and aredescribed, for example, in reference [52]. Examples of suitable carriersinclude lactose, starch, glucose, methyl cellulose, magnesium stearate,mannitol, sorbitol and the like. Examples of suitable diluents includeethanol, glycerol and water. The choice of pharmaceutical carrier,excipient or diluent can be selected with regard to the intended routeof administration and standard pharmaceutical practice. Thepharmaceutical compositions may comprise as, or in addition to, thecarrier, excipient or diluent any suitable binder(s), lubricant(s),suspending agent(s), coating agent(s), solubilising agent(s). Examplesof suitable binders include starch, gelatin, natural sugars such asglucose, anhydrous lactose, free-flow lactose, beta-lactose, cornsweeteners, natural and synthetic gums, such as acacia, tragacanth orsodium alginate, carboxymethyl cellulose and polyethylene glycol.Examples of suitable lubricants include sodium oleate, sodium stearate,magnesium stearate, sodium benzoate, sodium acetate, sodium chloride andthe like. Preservatives, stabilizers, dyes and even flavouring agentsmay be provided in the pharmaceutical composition. Examples ofpreservatives include sodium benzoate, sorbic acid and esters ofp-hydroxybenzoic acid. Antioxidants and suspending agents may be alsoused.

The compositions of the invention may be formulated as a food product.For example, a food product may provide nutritional benefit in additionto the therapeutic effect of the invention, such as in a nutritionalsupplement. Similarly, a food product may be formulated to enhance thetaste of the composition of the invention or to make the compositionmore attractive to consume by being more similar to a common food item,rather than to a pharmaceutical composition. In certain embodiments, thecomposition of the invention is formulated as a milk-based product. Theterm “milk-based product” means any liquid or semi-solid milk- orwhey-based product having a varying fat content. The milk-based productcan be, e.g., cow's milk, goat's milk, sheep's milk, skimmed milk, wholemilk, milk recombined from powdered milk and whey without anyprocessing, or a processed product, such as yoghurt, curdled milk, curd,sour milk, sour whole milk, butter milk and other sour milk products.Another important group includes milk beverages, such as whey beverages,fermented milks, condensed milks, infant or baby milks; flavoured milks,ice cream; milk-containing food such as sweets.

In certain embodiments, the compositions of the invention contain asingle bacterial strain or species and do not contain any otherbacterial strains or species. Such compositions may comprise only deminimis or biologically irrelevant amounts of other bacterial strains orspecies. Such compositions may be a culture that is substantially freefrom other species of organism.

The compositions for use in accordance with the invention may or may notrequire marketing approval.

In some cases, the lyophilised bacterial strain is reconstituted priorto administration. In some cases, the reconstitution is by use of adiluent described herein.

The compositions of the invention can comprise pharmaceuticallyacceptable excipients, diluents or carriers.

In certain embodiments, the invention provides a pharmaceuticalcomposition comprising: a bacterial strain of the invention; and apharmaceutically acceptable excipient, carrier or diluent; wherein thebacterial strain is in an amount sufficient to treat a disorder whenadministered to a subject in need thereof; and wherein the disorder isselected from the group consisting of asthma, allergic asthma,neutrophilic asthma, osteoarthritis, psoriatic arthritis, juvenileidiopathic arthritis, neuromyelitis optica (Devic's disease), ankylosingspondylitis, spondyloarthritis, systemic lupus erythematosus, celiacdisease, chronic obstructive pulmonary disease (COPD), cancer, breastcancer, colon cancer, lung cancer, ovarian cancer, uveitis, scleritis,vasculitis, Behcet's disease, atherosclerosis, atopic dermatitis,emphysema, periodontitis, allergic rhinitis, and allograft rejection.

In certain embodiments, the invention provides pharmaceuticalcomposition comprising: a bacterial strain of the invention; and apharmaceutically acceptable excipient, carrier or diluent; wherein thebacterial strain is in an amount sufficient to treat or prevent adisease or condition mediated by IL-17 or the Th17 pathway. In preferredembodiments, said disease or condition is selected from the groupconsisting of rheumatoid arthritis, multiple sclerosis, psoriasis,inflammatory bowel disease, Crohn's disease, ulcerative colitis, celiacdisease, asthma, allergic asthma, neutrophilic asthma, osteoarthritis,psoriatic arthritis, juvenile idiopathic arthritis, neuromyelitis optica(Devic's disease), ankylosing spondylitis, spondyloarthritis, systemiclupus erythematosus, chronic obstructive pulmonary disease (COPD),cancer, breast cancer, colon cancer, lung cancer, ovarian cancer,uveitis, scleritis, vasculitis, Behcet's disease, atherosclerosis,atopic dermatitis, emphysema, periodontitis, allergic rhinitis, andallograft rejection.

In certain embodiments, the invention provides the above pharmaceuticalcomposition, wherein the amount of the bacterial strain is from about1×10³ to about 1×10¹¹ colony forming units per gram with respect to aweight of the composition.

In certain embodiments, the invention provides the above pharmaceuticalcomposition, wherein the composition is administered at a dose of 1 g, 3g, 5 g or 10 g.

In certain embodiments, the invention provides the above pharmaceuticalcomposition, wherein the composition is administered by a methodselected from the group consisting of oral, rectal, subcutaneous, nasal,buccal, and sublingual.

In certain embodiments, the invention provides the above pharmaceuticalcomposition, comprising a carrier selected from the group consisting oflactose, starch, glucose, methyl cellulose, magnesium stearate, mannitoland sorbitol.

In certain embodiments, the invention provides the above pharmaceuticalcomposition, comprising a diluent selected from the group consisting ofethanol, glycerol and water.

In certain embodiments, the invention provides the above pharmaceuticalcomposition, comprising an excipient selected from the group consistingof starch, gelatin, glucose, anhydrous lactose, free-flow lactose,beta-lactose, corn sweetener, acacia, tragacanth, sodium alginate,carboxymethyl cellulose, polyethylene glycol, sodium oleate, sodiumstearate, magnesium stearate, sodium benzoate, sodium acetate and sodiumchloride.

In certain embodiments, the invention provides the above pharmaceuticalcomposition, further comprising at least one of a preservative, anantioxidant and a stabilizer.

In certain embodiments, the invention provides the above pharmaceuticalcomposition, comprising a preservative selected from the groupconsisting of sodium benzoate, sorbic acid and esters ofp-hydroxybenzoic acid.

In certain embodiments, the invention provides the above pharmaceuticalcomposition, wherein said bacterial strain is lyophilised.

In certain embodiments, the invention provides the above pharmaceuticalcomposition, wherein when the composition is stored in a sealedcontainer at about 4.0 or about 25.0 and the container is placed in anatmosphere having 50% relative humidity, at least 80% of the bacterialstrain as measured in colony forming units, remains after a period of atleast about: 1 month, 3 months, 6 months, 1 year, 1.5 years, 2 years,2.5 years or 3 years.

Culturing Methods

The bacterial strains for use in the present invention can be culturedusing standard microbiology techniques as detailed in, for example,references [53-55].

Bacterial strains of the genus Bacteroides may be cultured using amethod such as that outlined below, which may provide good growth andreliability. This method is particularly useful for culturing strains ofthe species Bacteroides coprocola.

Bacterial strains of the genus Bacteroides, and in particular of thespecies Bacteroides coprocola, may be cultured by using a liquid stockto inoculate the plate (or a larger liquid culture), rather than ascrape of frozen stock as may generally be used (see, for example,reference [56]). The establishment of mature colonies is reliable andquick if frozen stocks of Bacteroides strains are thawed and used as aliquid culture.

A method suitable for culturing a bacterial strain of the genusBacteroides, may comprise:

-   -   (a) providing a frozen stock of the bacterial strain;    -   (b) thawing the frozen stock to provide a liquid stock;    -   (c) adding the liquid stock to a solid or liquid medium; and    -   (d) incubating the solid or liquid media to provide a culture of        the bacterial strain.

This method is particularly useful for culturing bacterial strains ofBacteroides coprocola.

The frozen stock may be a frozen glycerol stock. The solid or liquidmedium may be YCFA agar or YCFA medium. YCFA medium may include (per 100ml, approximate values): Casitone (1.0 g), yeast extract (0.25 g),NaHCO₃ (0.4 g), cysteine (0.1 g), K₂HPO₄ (0.045 g), KH₂PO₄ (0.045 g),NaCl (0.09 g), (NH₄)₂SO₄ (0.09 g), MgSO₄.7H₂O (0.009 g), CaCl₂ (0.009g), resazurin (0.1 mg), hemin (1 mg), biotin (1 μg), cobalamin (1 μg),p-aminobenzoic acid (3 μg), folic acid (5 μg), and pyridoxamine (15 μg).

The incubating in step (d) may be performed for at least 36 hours, suchas 48 or 72 hours. The incubating in step (d) may be performed in ananaerobic environment, such as an anaerobic hood. The culture providedin step (d) may be used to subculture a larger culture of the bacterialstrain. Such subculturing allows greater amounts of bacteria to beprepared and may be useful for providing compositions of the inventionat a commercial scale.

The thawing in step (b) may be performed at room temperature, or by handwarming.

The amount of liquid stock added in step (c) may be between 300 μland 5ml, such 500 μl. The stock may be 20% glycerol stock.

The above method may be used for preparing a pharmaceutical compositionor a food product, in which case the method may comprise additionalsteps of isolating the bacterial strain, optionally lyophilising thebacterial strain, and combining the bacterial strain with one or morepharmaceutically acceptable excipients or carriers, or one or more foodsubstances. This pharmaceutical composition or food product prepared bythe method may be used in a method of treating or preventing a diseaseor condition mediated by IL-17 or the Th17 pathway.

An exemplary method of culturing bacterial strains of the genusBacteroides, and in particular of the species Bacteroides coprocola, maycomprise:

-   -   1. 500 μl of 20% glycerol stock is plated onto YCFA agar.    -   2. Plates are left in the anaerobic hood for 48/72 hours to        generate mature colonies.    -   3. The bacteria are cultured in 10 ml volumes, whether this be        from a single colony or a 1% liquid subculture.    -   4. Plates with colonies are only used for ⅔ days after mature        colony morphology is achieved.        Bacterial Strains for Use in Vaccine Compositions

The inventors have identified that the bacterial strains of theinvention are useful for treating or preventing diseases or conditionsmediated by IL-17 or the Th17 pathway. This is likely to be a result ofthe effect that the bacterial strains of the invention have on the hostimmune system. Therefore, the compositions of the invention may also beuseful for preventing diseases or conditions mediated by IL-17 or theTh17 pathway, when administered as vaccine compositions. In certain suchembodiments, the bacterial strains of the invention may be killed,inactivated or attenuated. In certain such embodiments, the compositionsmay comprise a vaccine adjuvant. In certain embodiments, thecompositions are for administration via injection, such as viasubcutaneous injection.

General

The practice of the present invention will employ, unless otherwiseindicated, conventional methods of chemistry, biochemistry, molecularbiology, immunology and pharmacology, within the skill of the art. Suchtechniques are explained fully in the literature. See, e.g., references[57] and [58-64], etc.

The term “comprising” encompasses “including” as well as “consisting”e.g. a composition “comprising” X may consist exclusively of X or mayinclude something additional e.g. X+Y.

The term “about” in relation to a numerical value x is optional andmeans, for example, x±10%.

The word “substantially” does not exclude “completely” e.g. acomposition which is “substantially free” from Y may be completely freefrom Y. Where necessary, the word “substantially” may be omitted fromthe definition of the invention.

References to a percentage sequence identity between two nucleotidesequences means that, when aligned, that percentage of nucleotides arethe same in comparing the two sequences. This alignment and the percenthomology or sequence identity can be determined using software programsknown in the art, for example those described in section 7.7.18 of ref.[65]. A preferred alignment is determined by the Smith-Waterman homologysearch algorithm using an affine gap search with a gap open penalty of12 and a gap extension penalty of 2, BLOSUM matrix of 62. TheSmith-Waterman homology search algorithm is disclosed in ref. [66].

Unless specifically stated, a process or method comprising numeroussteps may comprise additional steps at the beginning or end of themethod, or may comprise additional intervening steps. Also, steps may becombined, omitted or performed in an alternative order, if appropriate.

Various embodiments of the invention are described herein. It will beappreciated that the features specified in each embodiment may becombined with other specified features, to provide further embodiments.In particular, embodiments highlighted herein as being suitable, typicalor preferred may be combined with each other (except when they aremutually exclusive).

MODES FOR CARRYING OUT THE INVENTION Example 1 Efficacy of BacterialInocula in a Mouse Model of House Dust Mite-Induced Asthma

Summary

Mice were administered with compositions comprising bacterial strainsaccording to the invention and were subsequently challenged with housedust mite (HDM) extract to elicit an allergic inflammatory response. Theinflammatory response to HDM includes eosinophilic and neutrophiliccomponents, is mediated by IL-17 and the Th17 pathway, and is a modelfor asthma. The magnitude and characteristics of the inflammatoryresponse exhibited by mice treated with compositions of the inventionwere compared to control groups. The compositions of the invention werefound to alleviate the inflammatory response, and to reduce recruitmentof eosinophils and neutrophils, indicating that they may be useful fortreating IL-17- and Th17-mediated conditions such as eosinophilia,neutrophilia and asthma.

Strain

675: Bacteroides coprocola

Study Design

Groups:

1. Negative control group. Treatment with vehicle control (per oral).

3. Treatment with therapeutic bacteria inoculum strain 675 (per oral).

7. Positive control group. Treatment with Dexamethasone (i.p.).

8. Untreated Control Group.

Number of mice per group=5

Day −14 to day 13: Daily administration of vehicle control per oral(Group 1).

Day −14 to day 13: Daily administration of therapeutic bacteria inoculumper oral (Group 2-6).

Day 0, 2, 4, 7, 9, 11 Administration of 15 ug HDM (house dust miteextract—Catalogue number: XPB70D3A25, Lot number: 231897, GreerLaboratories, Lenoir, N.C., USA) in a volume of 30 ul PBS per nasal(Group 1-8).

Day 0, 2, 4, 7, 9, 11 Administration of Dexamethasone (i.p., 3 mg/kg,Sigma-Aldrich, Catalogue number D1159) (Group 7).

Day 14 Sacrifice of all animals for analysis.

Total number of mice=40.

Endpoints and Analysis

On day 14 animals were sacrificed by lethal intraperitoneal injectionwith pentabarbitol (Streuli Pharma AG, Uznach, Cat: 1170139A)immediately followed by a bronchoalveolar lavage (BAL).

Cells were isolated from the BAL (bronchoalveolar lavage) fluid anddifferential cell counts performed (200 cell counts/samples).

Material and Methods

Mice. Female 7 week old BALB/c mice were purchased from Charles RiverLaboratories and randomly allocated to cages totally 5 mice per cage(Ventilated cages sourced from Indulab AG, Gams, Switzerland Cage type:“The Sealsafe™—IVC cage. Product number 1248L). Cages were labeled withstudy number, group number and experimental starting date. Mice weremonitored weekly and acclimatized to facility for 7 days prior toinitiation of study (Study Day −14). Animals were 8 weeks old on StudyDay −14. Potable water and food were available ad libitum. Cageenrichment was present. Daily care of the animals was performedaccording to local authorization license number 2283.1 (issued andapproved by: Service de la consommation et des affaires vétérinaires duCanton de Vaud). Potable water and food were available ad libitum andrefreshed once daily. Cage enrichment was present. Animal welfareregulations were observed as given by official authorities ofSwitzerland under ordinance 455.163 of the FVO (Federal VeterinaryOffice) on laboratory animal husbandry, production of geneticallymodified animals, and methods of animal experimentation.

Culturing of bacteria inoculum. Within a sterile workstation, acryo-vial of bacteria was thawed by warming in gloved hand and ˜0.7 mlof contents injected into a Hungate tube (Cat Number, 1020471,Glasgeratebau Ochs, Bovenden-Lenglern, Germany), containing 8 ml ofanaerobic YCFA. Two tubes per strain were usually prepared. The Hungatetubes were then incubated (static) at 37° C. for up to 24-26 hours (forstrain 675).

Since the bacterial ODs of inoculum strain 675 was found to be variable,3 different culturing approaches were performed each day. 2 vials werecultured as described above and a third sample was cultured utilizing a400 ul aliquot from the prior day's culture for seeding. On 4 treatmentdays the latter approach was utilized, because of poor growth from thefrozen stock. Of note, this approach resulted in robust growth ofbacterial strain 675 on all occasions.

Culturing of vehicle control. A Hungate tube containing 8 ml ofanaerobic YCFA was incubated (static) at 37° C. for 16 h.

Administration of bacteria inoculum or vehicle control. 400 ul ofcultured bacteria inoculum or vehicle control were administered per dayper oral gavage.

Intranasal sensitization. Mice were anesthetized by i.p. injection with9.75 mg xylasol and 48.75 mg ketasol per kg (Dr. E. Graeub AG, Bern,Switzerland) and administered with 15 ug of HDM (Catalogue number:XPB70D3A25, Lot number: 231897, Greer Laboratories, Lenoir, N.C., USA)in a volume of 30 ul PBS per nasal.

Preparation and administration of positive control compoundDexamethasone. Dexamethasone 21-phosphate disodium salt (Sigma-Aldrich,Catalogue number D1159, Lot N° SLBD.1030V) was solved in H₂O andadministered to the animals in a dose of 3 mg/kg in a volume of 200 ulper oral at days indicated in study protocol above.

Terminal procedure. On day 14 animals were sacrificed by lethal i.p.injection with pentabarbitol (Streuli Pharma AG, Uznach, Cat: 1170139A)immediately followed by bronchoalveolar lavage (BAL) in 500 ul ofsaline.

Measurement of cellular infiltrates into BAL. Cells were isolated fromthe BAL fluid and differential cell counts were performed based uponstandard morphological and cytochemical criteria.

Graphs and statistical analysis. All graphs were generated with GraphpadPrism Version 6 and a one-way ANOVA was applied. Results from thestatistical analysis were provided with the individual data tables.Error bars represent Standard Error of the Mean (SEM).

Results and Analysis

The results of the experiments are shown in FIGS. 1-9.

No morbidity or mortality was noted in the mice treated with thebacteria or the vehicle. The two controls, vehicle treatment (negativecontrol) and the dexamethasone treatment (positive control) behaved asexpected, with impaired eosinophilia and neutrophilia noted followingdexamethasone treatment.

The most important results of this experiment are displayed in FIGS. 6and 7, which report on the total number and percentage of neutrophilsdetected in bronchiolar lavage following challenge with HDM. Strain 675reduced total neutrophils and the proportion of neutrophils in BALrelative to the vehicle-only control.

Example 2 Efficacy of Bacterial Inocula in a Mouse Model of SevereNeutrophilic Asthma

Summary

Mice were administered with compositions comprising bacterial strainsaccording to the invention and were subsequently sensitised withsubcutaneous administrations of house dust mite (HDM) extract andchallenged with an intranasal administration of HDM in order to modelthe inflammatory response of severe neutrophilic asthma. The magnitudeand characteristics of the inflammatory response exhibited by micetreated with compositions of the invention were compared to controlgroups. The compositions of the invention were found to alleviate theinflammatory response, and in particular to reduce recruitment ofneutrophils, in a manner comparable to the positive control comprisingadministrations of anti-IL-17 antibodies. The data therefore indicatethat the compositions of the invention may be useful for treating IL-17-and Th17-mediated conditions such as neutrophilia and asthma.

Strain

675: Bacteroides coprocola

Study Design

Groups:

1. Negative control group. Treatment with vehicle control (per oral).

3. Treatment with therapeutic bacteria inoculum strain 675 (per oral).

7. Positive control group. Treatment anti-IL-17 (i.p.).

8. Untreated Control Group.

9: Healthy mice (baseline).

Number of mice per group (Group 1-8)=5

Day −14 to day 17: Daily administration of vehicle control per oral(Group 1).

Day −14 to day 17: Daily administration of therapeutic bacteria inoculumper oral (Group 2-6).

Day 0: Sensitization with HDM in CFA (s.c.) (Group 1-8).

Day 7: Sensitization with HDM in CFA (s.c.) (Group 1-8).

Day 13, 15, 17: Administration of anti IL-17 neutralizing antibody peri.p. (Group 7).

Day 14, 15, 16, 17: Challenge with HDM in 30 ul PBS per nasal (Group1-8).

Day 18: Sacrifice of all animals for analysis.

Endpoints and Analysis:

On day 14 animals were sacrificed by lethal intraperitoneal injectionwith pentabarbitol (Streuli Pharma AG, Uznach, Cat: 1170139A)immediately followed by a bronchoalveolar lavage (BAL). Cells wereisolated from the BAL fluid and differential cell counts performed (200cell counts/samples).

Material and Methods

Mice. Female 7 week old C57BL/6 mice were purchased from Charles RiverLaboratories and randomly allocated to cages totally 5 mice per cage(Ventilated cages sourced from Indulab AG, Gams, Switzerland Cage type:“The Sealsafe™—IVC cage. Product number 1248L). Cages were labelled withstudy number, group number and experimental starting date. Mice weremonitored weekly and acclimatized to facility for 7 days prior toinitiation of study (Study Day −14). Animals were 8 weeks old on StudyDay −14. Potable water and food were available ad libitum. Cageenrichment was present. Daily care of the animals was performedaccording to local authorization license number 2283.1 (issued andapproved by: Service de la consommation et des affaires vétérinaires duCanton de Vaud). Potable water and food were available ad libitum andrefreshed once daily. Cage enrichment was present. Animal welfareregulations were observed as given by official authorities ofSwitzerland under ordinance 455.163 of the FVO (Federal VeterinaryOffice) on laboratory animal husbandry, production of geneticallymodified animals, and methods of animal experimentation.

Culturing of bacteria inoculum. Within a sterile workstation, acryo-vial of bacteria was thawed by warming in gloved hand and ˜0.7 mlof contents injected into a Hungate tube (Cat Number, 1020471,Glasgeratebau Ochs, Bovenden-Lenglern, Germany), containing 8 ml ofanaerobic YCFA. Two tubes per strain were usually prepared. The Hungatetubes were then incubated (static) at 37° C. for up to 24-26 hours (forstrain 675).

Since the bacterial ODs of inoculum strain 675 were variable, 3different culturing approaches were performed each day. 2 vials werecultured as described above and a third sample was cultured utilizing a400u1 aliquot from the prior day's culture for seeding. On 4 treatmentdays the latter approach was utilized, because of poor growth from thefrozen stock. Of note, this approach resulted in robust growth ofbacterial strain 675 on all occasions.

Culturing of vehicle control. A Hungate tube containing 8 ml ofanaerobic YCFA was incubated (static) at 37° C. for 16 h.

Administration of bacteria inoculum or vehicle control. 400 ul ofcultured bacteria inoculum or vehicle control were administered per dayper oral gavage.

HDM sensitization. 50 μg of HDM (Catalogue number: XPB70D3A25, Lotnumber: 231897, Greer Laboratories, Lenoir, N.C., USA) in PBS wasemulsified in equal volume of complete Freund's adjuvant (CFA ChondrexInc. Washington, USA) and administered subcutaneously in a volume of 200μl, twice over two weeks on opposite flanks. A week after the secondimmunization, mice were anesthetized by i.p. injection with 9.75 mgxylasol and 48.75 mg ketasol per kg (Dr. E. Graeub AG, Bern,Switzerland) and then given intranasal challenges of 15 μg of HDM in avolume of 30 ul PBS on 4 consecutive days. Analysis was performed oneday after the final challenge.

Preparation and administration of positive control compound anti mouseIL-17 antibody. Anti-IL-17 neutralizing antibody was sourced from Bio XCell and was stored at 4° C. (Clone 17F3, Cat. Number BE0173, Bio XCell) and administered per i.p. at a dose of 12.5 mg/kg at daysindicated in study protocol above.

Terminal procedure. On day 18 animals were sacrificed by lethal i.p.injection with pentabarbitol (Streuli Pharma AG, Uznach, Cat: 1170139A)immediately followed by bronchoalveolar lavage (BAL) in 500 ul ofsaline.

Measurement of cellular infiltrates into BAL. Cells were isolated fromthe BAL fluid and differential cell counts were performed based uponstandard morphological and cytochemical criteria.

Graphs and statistical analysis. All graphs were generated with GraphpadPrism Version 6 and a one-way ANOVA was applied. Results from thestatistical analysis are provided with the individual data tables. Errorbars represent Standard Error of the Mean (SEM).

Results and Analysis

The results of the experiment are shown in FIGS. 10-18.

No morbidity or mortality was noted in the mice treated with thebacteria or the vehicle. As shown in FIGS. 15 and 16, strain 675exhibited a strong effect and reduced total neutrophil numbers relativeto the negative controls. In addition, strain 675 reduced eosinophilnumbers relative to the controls, as shown in FIGS. 11 and 12.

Example 3 Efficacy of Bacterial Inocula to Treat Arthritis in a Type IICollagen-Induced Arthritis Mouse Model

Materials and Methods

Strain

675: Bacteroides coprocola

Bacterial Cultures

Bacterial cultures were grown up for administration in an anaerobicworkstation (Don Whitley Scientific) according to the growth schemebelow.

-   -   Mon/Weds/Fri: Transfer glycerol stock to ice and streak a YCFA        plate from glycerol stock. Use plate as follows for a maximum of        3 days.    -   Daily PM: Pick single colony of each strain from plate cultures,        transfer to 8 ml hungate tube containing YCFA overnight (ON1)    -   AM: Subculture 80 ul (1%) ON1 into fresh 8 ml tube (DC1). Use        ON1 culture for AM oral gavage.    -   PM: Use DC1 culture for PM oral gavage.

Bacterial strain #675 was grown using glycerol stocks.

Glycerol stocks were stored at −80° C. Three times per week, glycerolstocks were thawed at room temperature and streaked on YCFA plates. Anew glycerol aliquot was used on each occasion. Bacteria were allowed togrow on a given plate for up to 72 hours.

Solutions to be administered to the animals were prepared twice dailywith an eight hour interval for morning (AM) and afternoon (PM)treatments. A bacterial colony was picked from the streaked plate andtransferred into a tube containing YCFA media. Bacterial strain #675 wasallowed to grow for 16 hours before AM administrations. Bacteria weresub-cultured at 1% into YCFA media for PM administrations. OD valueswere recorded for each strain after morning and afternoon treatmentpreparations.

Type II Collagen-Induced Arthritis Mouse Model

Adult male DBA/1 mice were randomly allocated to experimental groups andallowed to acclimatise for two weeks. On Day 0, animals wereadministered by subcutaneous injection with 100 microliters of anemulsion containing 100 micrograms of type II collagen (CII) inincomplete's Freund's adjuvant supplemented with 4 mg/ml Mycobacteriumtuberculosis H37Ra. On Day 21, animals were administered by subcutaneousinjection with a booster emulsion containing 100 μg of type II collagenin incomplete Freund's adjuvant.

Treatments were given according to the administration schedule below.From Day −14 until the end of the experiment on Day 45, animals wereweighed three times per week. From Day 21 until the end of theexperiment, animals were scored three times per week for clinical signsof arthritis to include swelling of the hind- and front paws,radio-carpal (wrist) joints and tibio-tarsal (ankle) joints.

On Day 45 mice were culled and terminal blood samples were taken forcytokine analysis.

On Day −14, Day 0 and Day 45, faecal samples were collected formicrobiological analysis, immediately snap-frozen and stored at −80° C.

The collagen-induced arthritis (CIA) mouse model is a well-establishedmouse model for rheumatoid arthritis [67]. Immunisation with CII causesa pathogenesis that includes several important pathological features ofrheumatoid arthritis, including synovial hyperplasia, mononuclear cellinfiltration and cartilage degradation. Significantly, the developmentof CIA is mediated by Th17 cells through secretion of IL-17A [68]. Theimmune response underlying the arthritis model is enhanced by the use ofFreund's adjuvant supplemented with Mycobacterium tuberculosis.

On Day 21, spleens were collected from three satellite animals in eachgroup. Cells were cultured for 72 hours in the presence or absence oftype II collagen. Cytokines, including TNF-α, IL-6, IFN-γ, IL-4, IL-10and IL-17, were quantified in the culture supernatants and in terminalserum by Luminex. Cell proliferation was quantified using a tritiatedthymidine incorporation method.

Treatment Groups and Dosages

All Groups were n=15 (n=12 for the main study group and n=3 forsatellite groups)

The vehicle used for the biotherapeutics was Yeastextract-Casitone-Fatty Acids (YCFA) medium.

Administration Disease Group Dose Route Regimen Induction 1 Vehicle 5ml/kg PO BID: Day 0: 4 Biotherapeutic #675 5 ml/kg Day-14*-EndCollagen/CFA, once, SC Day 21: Collagen/IFA, once, SC PO: oral gavage,SC: subcutaneous injection, BID: twice a day, CFA: complete Freund'sadjuvant. *Except Group 4 treated from Day 0.Bodyweights

From Day −14 until the end of the experiment, animals were weighed threetimes per week. Data were graphed (Mean±SEM).

Non-Specific Clinical Observations

From Day −14 until the end of the experiment, animals were checked dailyfor non-specific clinical signs to include abnormal posture (hunched),abnormal coat condition (piloerection) and abnormal activity levels(reduced or increased activity).

Clinical Observations

From Day 21 until the end of the experiment on Day 45, animals werescored three times per week for clinical signs of arthritis to includeswelling of the hind- and front paws, radio-carpal (wrist) joints andtibio-tarsal (ankle) joints. Each limb was scored using the followingscale: (0) normal, (1) slight swelling, (2) mild swelling, (3) moderateswelling and (4) severe swelling. A clinical score was calculated byadding each limb score. The maximum possible clinical score for ananimal was (16). Animals with a score equal to (12) on two consecutiveoccasions and animals with a score greater than (12) on any one occasionwere culled. Data were graphed (Mean±SEM).

Cell Proliferation Analysis

On Day 21, three satellite animals per group were culled and spleenswere dissected out. Spleen cells were cultured for 72 hours in presenceor absence of type II Collagen. After 72 hours, cells were pulsedovernight in the presence of tritiated thymidine. Cell proliferation wasquantified by measuring thymidine incorporation. Data were graphed(Mean±SEM). Supernatants were taken and tested for the presence of keycytokines.

Cytokine Analysis

Terminal supernatants from the spleen cell cultures were tested in orderto quantitate TNF-α, IL-6, IFN-Δ, IL-4, IL-10 and IL-17 by Luminex. Datawere graphed (Mean±SEM).

Microbiological Analysis

On Day −14, Day 0 and Day 45, faecal samples were collected from eachanimal, immediately snap-frozen, and stored at −80° C. Caeca (includingcontent) were immediately snap-frozen and stored at −80° C. A bacterialidentification test was performed daily by plating the bacteria.

Histopathology

At the end of the experiment, hind paws were stored in tissue fixative.Samples were transferred into decalcification solution. Tissue sampleswere processed, sectioned and stained with Haematoxylin & Eosin.Sections were scored by a qualified histopathologist, blind to theexperimental design, for signs of arthritis to include inflammation,articular cartilage damage and damage to the underlying metaphysealbone. A detailed scoring system was used (see below). Data were graphed(Mean±SEM). Raw and analysed data were provided as well asrepresentative pictures.

TABLE 1 Histopathology Scoring System Grade Description Inflammation 0Normal joint 1 Mild synovial hyperplasia with inflammation dominated byneutrophils. Low numbers of neutrophils and macrophages in joint space.2 Synovial hyperplasia with moderate to marked inflammation involvingboth neutrophils and macrophages. Neutrophils and macrophages in jointspace; may be some necrotic tissue debris. 3 Synovial hyperplasia withmarked inflammation involving both neutrophils and macrophages. Loss ofsynoviocyte lining. Inflam- mation may extend from synovium tosurrounding tissue includ- ing muscle. Numerous neutrophils andmacrophages in joint space, together with significant necrotic tissuedebris. Articular cartilage damage 0 Normal joint 1 Articular cartilageshows only mild degenerative change. Early pannus formation may bepresent peripherally. 2 Articular cartilage shows moderate degenerativechange and focal loss. Pannus formation is present focally. 3Significant disruption and loss of articular cartilage with exten- sivepannus formation. Damage to the underlying metaphyseal bone 0 Normaljoint 1 No change to underlying metaphyseal bone. 2 May be focalnecrosis or fibrosis of metaphyseal bone. 3 Disruption or collapse ofmetaphyseal bone. Extensive inflamma- tion, necrosis or fibrosisextending to medullary space of the metaphysis.Results and AnalysisSurvival and Non-Specific Clinical Observations

Some animals were culled prior to the scheduled end of the study due tothe severity of the clinical signs of arthritis or due to the severityof the non-specific clinical observations.

One animal in Group 1 was culled (vehicle-treated, animal arrived fromthe supplier with broken leg).

Eleven animals were culled due to the severity of the clinical signs ofarthritis: five animals in Group 1 (vehicle-treated), and six animals inGroup 4 (biotherapeutic #675-treated).

Five animals were culled due to the severity of the non-specificclinical signs including abnormal posture (hunched), abnormal coatcondition (piloerection), abnormal activity levels (reduced activity):three animals in Group 1 (vehicle-treated) and two animals in Group 4(biotherapeutic #675-treated).

Bodyweights

Bodyweight data recorded from Day −14 until Day 0 and expressed as apercentage of the initial (Day −14) bodyweights were analysed by two-wayANOVA followed by Dunnett's post-test for multiple comparisons with Day−14 then for multiple comparison with the vehicle-treated group. Thedata are presented in FIG. 19. Data from animals culled prior to thescheduled end of the experiment were excluded from the analyses.

When compared to Day −14, twice daily administrations by oral gavageinduced a significant bodyweight loss in the vehicle-treated group onDay −9 and Day −7.

Group 4 (untreated until Day 0 then biotherapeutic #675-treated)bodyweights were significantly higher than in the vehicle-treated groupfrom Day −11 until Day −1 (p<0.0001 except Day −4 where p<0.05).

The bodyweights measured between Day −14 and Day −1 in thebiotherapeutic-treated groups did not differ from the bodyweightsmeasured in the vehicle-treated group on any given day.

Bodyweight data recorded from Day 0 until Day 28 and expressed as apercentage of the initial (Day 0) bodyweights were analysed by two-wayANOVA followed by Dunnett's post-test for multiple comparisons with Day0 in the Vehicle group then for multiple comparison with thevehicle-treated group. The data are presented in FIG. 20. Data fromanimals culled prior to the scheduled end of the experiment and fromSatellite animals were excluded from the analyses. Day 28, Day 35 andDay 42 data were further analysed by one-way ANOVA followed by Dunnett'spost-test for multiple comparisons to the vehicle-treated group.

The onset of clinical signs of arthritis was associated with asignificant bodyweight loss on Day 26 and Day 28 (p<0.0001) whencompared to Day 0 in the vehicle-treated group.

When compared to the vehicle-treated group, the bodyweights weresignificantly higher in Group 4 (biotherapeutic #675-treated) on Day 3,Day 5, Day 10 (p<0.05) and on Day 26 (p<0.001).

When analysing the data by one-way ANOVA, the bodyweights weresignificantly higher in Group 4 (biotherapeutic #675-treated) whencompared to the vehicle-treated group on Day 28 (p<0.01). There was nosignificant difference between experimental groups on Day 35 or Day 42.

Clinical Observations

Clinical score data were analysed by two-way ANOVA followed by Dunnett'spost-test for multiple comparisons between days in the vehicle-treatedgroup then for multiple comparisons between experimental groups and thevehicle-treated group each day. The data are presented in FIG. 21. Datarecorded from animals culled prior to the end of the experiment wereexcluded from the analysis. When animals were culled due to the severityof the clinical signs of arthritis, the last recorded score was reportedfor the following days and used in the statistical analyses.

A significant increase of the clinical scores was observed in thevehicle-treated group from Day 28 until Day 45 (p<0.0001) when comparedto Day 21.

Biotherapeutic #675 did not reduce the clinical scores when compared tothe vehicle-treated group. Animals in this group were immunised at adifferent time when compared to other experimental groups, which mayexplain the higher clinical scores observed.

Cell Proliferation Analysis

To validate the assay, splenocytes were cultured in the presence ofsoluble anti-CD3 and anti-CD28 (anti-CD3/CD28) as positive controlstimuli to confirm the proliferative potential of the cells.

Strong proliferative responses to anti-CD3/CD28 were seen in allexperimental groups, showing cells were healthy, viable and able torespond to activation signals.

To test the proliferative response in presence of Collagen II (CII),splenocytes were cultured in the presence of CII at 50 mg/ml. Splenocyteproliferative response to CII were analysed by two-way

ANOVA followed by Sydak's post-test for multiple comparisons betweenunstimulated and CII-stimulated splenocytes and one-way ANOVA followedby Dunnett's post-test for comparison of CII-stimulated response indifferent experimental groups with the vehicle-treated group. The dataare presented in FIG. 22.

CII induced a highly significant increase of ³H-thymidine incorporation(cpm) when compared to the unstimulated splenocytes in thevehicle-treated group (p<0.0001).

Splenocyte proliferation for group treated with biotherapeutic #675 wasset up on a different day, therefore comparison with the vehicle-treatedgroup was not preformed, although a notable reduction was observed.

Cytokine Levels in Tissue Culture Supernatants

Levels of each cytokine were measured in tissue culture supernatantsderived from anti-CD3/CD28 stimulated cultures by luminex analysis.These showed robust responses for all cytokines measured (mean levels invehicle group were as follows: IL-4=6,406 pg/ml; IL-6=306 pg/ml;IL-10=10,987 pg/ml; IL-17A=11,447 pg/ml; IFN-γ=15,581 pg/ml; TNF-α=76pg/ml).

The following sections summarise the data obtained from the CollagenII-stimulated cultures. Where applicable, statistical analyses of thedifferences between cytokine levels in supernatants of unstimulated andCII-stimulated splenocytes were conducted using two-way ANOVA followedby Sidak's post-test for multiple comparisons, while one-way ANOVAfollowed by Dunnett's post-test was used for comparison ofCII-stimulated response in biotherapeutic-treated groups with thevehicle-treated group. There was no significant difference in cytokinelevels between the groups in both cases. This is likely due to the smallsample size used (n=3).

In order to more accurately present the distribution of the data for thecytokines with substantial spread of the data, these are presented asscatter plots.

The group means of IL-4 in tissue culture supernatants after stimulationwith CII were <5 pg/ml. These are not considered biologicallysignificant and not included here. The group means of TNF-α in tissueculture supernatants after stimulation with collagen were below limit ofquantitation.

Supernatant Levels of IFN-γ (FIG. 23)

Along with IL-17, IFN-γ is the major cytokine driving disease in the CIAmodel. The scatter plot in FIG. 23 demonstrates IFN-γ levels after CIIstimulation, with group median being higher for the Vehicle-treatedgroup compared to the biotherapeutic (see FIG. 27).

Supernatant Levels of IL-17A (FIG. 24)

Levels of IL-17A were 50 pg/ml in CII-stimulated cultures for theVehicle-treated group. The levels of this cytokine appeared to be lowerin the biotherapeutic group compared to the Vehicle-treated group (seeFIG. 27).

Supernatant Levels of IL-10 (FIG. 25)

Levels of IL-10 in Vehicle-treated group were 13 pg/ml and 2.1 pg/ml forCII-stimulated, and media control cultures, respectively. Higher levelsof IL-10 (which is an anti-inflammatory cytokine) for thevehicle-treated group may be expected because inflammation andpro-inflammatory cytokine induction could be accompanied by ananti-inflammatory feedback mechanism.

Supernatant Levels of IL-6 (FIG. 26)

Inflammatory cytokines such as IL-6 and TNF-α are not typically producedat high levels in anti-CII cultures. However, their levels may bealtered as a result of immune modulation. Levels of IL-6 inCII-stimulated cultures were modest, reaching 10 pg/ml. Although higherthan in media control cultures, these differences were too small toprovide rationale for performing statistical analyses.

Tissue Culture Supernatants in Group 4—Biotherapeutic #675 (FIG. 27)

Splenocyte cultures for this group were set up on a different day andare therefore separate from the Vehicle-treated group. Although directcomparisons may not be appropriate, it appears that treatment withBiotherapeutic #675 was effective at lowering IFN-γ, IL-17A and IL-6levels.

Microbiological Analysis

Bacterial growth was confirmed by measuring the optical density at 600nm using a spectrophotometer. Bacterial identity was confirmed bycomparing streaked plate pictures to reference pictures.

Following the improved bacterial preparation method, consistently highdoses of bacterial strain were administered from Day −2 and Day −3(except for strain #675 from Day 0) as indicated by the high OD valuesmeasured.

Faecal samples were collected and snap-frozen on Day −14, Day 0 and attermination.

Histopathology

The histopathology results are shown in FIGS. 66-71. As expected forthis model, intra-individual and inter-individual variability wasobserved in terms of the presence/absence of arthritis or the severityof change present.

The nature of the pathology was as expected for this model, withextensive mixed chronic-active inflammation of the synovium and bursaextending to involve the peri-articular soft tissues (muscle, adiposetissue, dermal collagen). In the most severely affected joints there wasarticular cartilage degeneration and loss with intra-articular debrisand inflammation and disruption of the joint and bone structure byfibrosis and inflammation.

The incidence of histopathological changes was: vehicle—80% (16/20);Biotherapeutic #675-83% (20/24). Biotherapeutic #675 was effective forreducing histopathological damage observed in hind limb joints andreducing joint inflammation scores, cartilage damage scores, bone damagescores and total histopathology scores (see FIG. 70), althoughstatistical comparisons with the vehicle group could not be performed.

Summary

Increased clinical scores were observed from Day 28 after the firstadministration of type II collagen, as expected in this model ofarthritis in DBA/1 mice. Biotherapeutic #675 was shown to be effectiveat treating arthritis in this model. Animals treated with biotherapeutic#675 were immunised at a different time when compared to thevehicle-treated group, which may explain the higher clinical scoresobserved. However, biotherapeutic #675 was effective for reducingpathological disease in the joints, as demonstrated in thehistopathological analysis.

Proliferative recall responses to Collagen II were seen in splenocytecultures from all experimental groups. Statistics were not performed forcultures of biotherapeutic #675, as these were established at adifferent time, but a reduction in collagen-specific response relativeto the control was observed.

Most of the T cell cytokines tested showed detectable increases betweenCollagen II-stimulated and media controls in the Vehicle-treated group.These increases were not as obvious in the biotherapeutic-treated group.This broadly supports the proliferative recall responses to Collagen IIdescribed above.

There was evidence of suppression of the Th1/Th17 axis, which is thepathogenic response in this model and in human RA. Correlation ofreduced levels of cytokines with reduced proliferation is suggestive ofimmune modulation. There was no evidence that this modulation resultedeither from enhanced levels of Th2 associated IL-4 or with increases inthe immune modulating cytokine, IL-10.

Example 4 Further Analysis of the Effect of Bacterial Inocula in theMouse Model of House Dust Mite-Induced Asthma

The mice tested in Example 1 were subjected to further analyses tofurther characterise the effect of the compositions of the invention onthe allergic asthma inflammatory response.

Materials and Methods

Blood withdrawal and serum preparation on day 14. Blood samples ofanimals were collected via cardiac puncture. Serum was isolated from theblood sample by centrifugation for 5 min at 14000 g and stored at −20°C.

Organ removal on day 14. Collection of the left lung lobe in formalinfor follow-on histological analysis. Collection of the right lung lobes(all remaining lobes) and removal of serum for snap freezing andfollow-on analysis. Remaining BAL fluid was snap frozen for follow-onanalysis.

Measurement of Antibody Levels in Serum and BAL Fluid

Total IgE and house-dust-mite (HDM) specific IgG1 antibody productionwere measured in the BAL and serum by ELISA assay.

Isolation of Lung and Histological Analysis

Left lung lobes were fixed in formalin followed by embedment inparaffin, sectioning, and staining with hematoxylin and eosin and PAS.Subsequent histological scoring was performed blinded as followed: Fiverandom fields of view per sample were scored for inflammation(peribronchial infiltration and perivascular infiltration) and mucusproduction. Inflammatory infiltration was scored with the followinggrading system:

0—normal

1—mild inflammatory infiltrates

2—moderate inflammatory infiltrates

3—marked inflammatory infiltrates

4—severe inflammatory infiltrates

5—very severe inflammatory infiltrates

In each field of view, airways were measured in size and mucus cellnumbers were quantified/um.

Measurement of Inflammatory Mediators in Lung Tissue

Right lung lobes (all remaining lobes) isolated for quantification ofinflammatory mediators were snap frozen for subsequent measurement ofCCL11, IFN-gamma, IL-1 alpha, IL-1 beta, IL-4, IL-5, IL-9, IL-17A,CXCL1, CCL3, CXCL2 and CCL5 by commercially available multiplex assay(Merck-Millipore). Analysis was performed according to themanufacturer's instructions.

Results and Analysis

The results of the experiments are shown in FIGS. 28-46.

In support of the findings described in Example 1, analysis of thecellular infiltrates in the lung tissue of mice treated with strain 675showed a notable and statistically significant reduction in meaninflammation score (see FIGS. 32 and 34).

Antibody levels in the BAL fluid and serum were analysed (see FIGS.28-31). No clear effect of the bacterial treatment on serum antibodylevels was observed. This may reflect a failure in the experiment,because the spread of data and the error bars for each treatment arelarge, and the positive and negative controls do not appear to havebehaved as would be expected. Also, the baseline serum antibody levelscould have masked any changes.

Similarly, no clear effect of the bacterial treatment on cytokine levelsin lung tissue was observed (see FIGS. 36-46). Again, this may reflect afailure in the experiment, because the spread of data and the error barsfor each treatment are large, and the positive and negative controls donot appear to have behaved as would be expected. It is also possiblethat the mechanism of action involved influences earlier cytokineresponses that were no longer detectable on day 4 post the final HDMairway challenge. Some care should be taken when interpreting thecytokine data in the current study, due to the variability in the levelsdetected. This variability could in part be explained by the fact thatthe lung tissue was separated for the different analyses, and thus onelung lobe might not have been fully representative or comparable to thesame lobe in other mice due to patchy distribution of the inflammation.

Example 5 Further Analysis of the Effect of Bacterial Inocula in theMouse Model of Severe Neutrophilic Asthma

The mice tested in Example 2 were subjected to further analyses tofurther characterise the effect of the compositions of the invention onthe neutrophilic response associated with severe asthma.

Materials and Methods

Organ removal on day 18. Collection of the left lung lobe in formalinfor follow-on histological analysis. Collection of the right lung lobes(all remaining lobes) and removal of serum for snap freezing andfollow-on analysis. Remaining BAL fluid was snap frozen for follow-onanalysis.

Measurement of inflammatory mediators in lung tissue (follow-onanalysis). Right lung lobes (all remaining lobes) isolated forquantification of inflammatory mediators were snap frozen for subsequentmeasurement of IFN-gamma, IL-1 alpha, IL-1 beta, CXCL1, CCL3, CXCL2,CCL5, IL-17A, TNF-alpha, IL-17F, IL-23 and IL-33 by commerciallyavailable multiplex assay (Merck-Millipore). Analysis was performedaccording to the manufacturer's instructions.

Measurement of antibody levels in serum and BAL fluid (follow-onanalysis). House-dust-mite (HDM) specific IgG1 and IgG2a antibodyproduction were measured in the BAL and serum by ELISA assay.

Isolation of lung and histological analysis (follow-on analysis). Leftlung lobes were fixed in formalin followed by embedment in paraffin,sectioning, and staining with hematoxylin and eosin and PAS. Subsequenthistological scoring was performed blinded as followed: Five randomfields of view per sample were scored for inflammation (peribronchialinfiltration and perivascular infiltration) and mucus production.Inflammatory infiltration was scored with the following grading system:

0—normal

1—mild inflammatory infiltrates

2—moderate inflammatory infiltrates

3—marked inflammatory infiltrates

4—severe inflammatory infiltrates

5—very severe inflammatory infiltrates

Results and Analysis

The results of the experiments are shown in FIGS. 47-64.

Further analysis of antibody levels revealed that the efficacy ofbacterial strain 675 was also reflected in reduced HDM-specific IgG1levels in the BAL fluid and serum (see FIGS. 47 and 49). Firmconclusions regarding an effect on IgG2a levels cannot be drawn.Overall, the data from the antibody analysis is suggestive of areduction related to an overall reduced inflammatory response, asopposed to a selective effect on antibody isotype switching.

Histological analysis supported the differential cell counts from theBAL fluid, showing a reduced cellular infiltrate in mice treated withStrain 675 (see FIGS. 51-53).

In relation to cytokine levels, as for Example 4, the spread of data andthe error bars for each treatment are large, and the positive andnegative controls do not appear to have behaved as necessarily would beexpected. It is also possible that the mechanism of action involvesinfluencing earlier cytokine responses that were no longer detectable onday 4 post the final HDM airway challenge. Some care should be takenwhen interpreting the cytokine data in the current study, due to thevariability in the levels detected. This variability could in part beexplained by the fact that the lung tissue was separated for thedifferent analyses, and thus one lung lobe might not have been fullyrepresentative or comparable to the same lobe in other mice due topatchy distribution of the inflammation. Despite this variability, aclear anti-inflammatory effect on cytokine levels for strain 675 wasshown, and the positive control anti-IL-17 Ab generally behaved asexpected.

With the above caveats, the data in FIGS. 56, 58, 59, 61 and 63 suggestthat treatment with the bacterial strains of the invention may achieve areduction in the levels of IL-1b, IFNγ, RANTES, MIP-1a and KC (the mouseorthologue of human IL-8), which may be indicative of a mechanism ofaction related to influences on chemokine release (and thus recruitmentof cells) by stromal or innate immune cells. These cytokines are part ofthe Th17 pathway. Taking this dataset together, a clear conclusion canbe drawn that Strain 675 was highly effective at protecting mice againstinflammation in this mouse model of severe neutrophilic asthma.

Example 6 Efficacy of Bacterial Inocula in a Mouse Model of MultipleSclerosis

Summary

Mice were administered with compositions comprising bacterial strainsaccording to the invention and the mice were subsequently immunised withmyelin oligodendrocyte glycoprotein to induce experimental autoimmuneencephalomyelitis (EAE). EAE is the most commonly used experimentalmodel for human multiple sclerosis. The compositions of the inventionwere found to have a striking effect on disease incidence and diseaseseverity.

Strain

675: bacteria deposited under accession number NCIMB 42408

Study Design

Groups:

1. Negative control group. Treatment with vehicle control (per oral).

3. Treatment with therapeutic bacteria inoculum strain 675 (per oral).

9. Positive control group. Treatment with Dexamethasone (i.p.).

10. Untreated Control Group.

Number of mice per group=10

Days −14 to day 27: Daily administration of vehicle control per oral(Group 1).

Days −14 to day 27: Daily administration of therapeutic bacteriainoculum per oral (Group 4).

Days 0-28: administration of Dexamethasone (i.p.) three times a week(Group 9)

Day 0: MOG35-55 (myelin oligodendrocyte glycoprotein—2 mg/ml) and CFA (2mg/ml MTB) were mixed 1:1 resulting in 1 mg/ml solutions. 100 μl of thepeptide-CFA mixture was injected subcutaneously into each hind leg.Administration of pertussis toxin intraperitoneally (300 ng).

Day 1: Administration of pertussis toxin intraperitoneally (300 ng).

Days 7-onwards: Measurement of disease incidence and weight three timesa week.

Endpoints and Analysis

Mice were analysed for disease incidence and disease severity threetimes a week. Scoring was performed blind. Disease severity was assessedusing a clinical score ranging from 0 to 5, with 5 indicating a deadmouse (see clinical scoring system below).

Monitoring

On the indicated days mice were weighed and observed for diseaseactivity score and disease incidence.

Disease activity score observations:

0—No obvious changes in motor function compared to non-immunized mice.

0.5—Tip of tail is limp.

1.0—Limp tail.

1.5—Limp tail and hind leg inhibition.

2.0—Limp tail and weakness of hind legs.

-   -   OR—There are obvious signs of head tilting when the walk is        observed. The balance is poor.

2.5—Limp tail and dragging of hind legs.

-   -   OR—There is a strong head tilt that causes the mouse to        occasionally fall over.

3.0—Limp tail and complete paralysis of hind legs.

3.5—Limp tail and complete paralysis of hind legs.

-   -   In addition to: Mouse is moving around the cage, but when placed        on its side, is unable to right itself.    -   Hind legs are together on one side of body.

4.0—Limp tail, complete hind leg and partial front leg paralysis.

-   -   Mouse is minimally moving around the cage but appears alert and        feeding

4.5—Complete hind and partial front leg paralysis, no movement aroundthe cage.

-   -   Mouse is immediately euthanized and removed from cage.

5.0 Mouse is euthanized due to severe paralysis.

When an animal has equal or greater disease activity score of 1, it isconsidered to have a positive disease incidence score.

Results

The results of the study are shown in FIGS. 72 and 73.

Disease induction in the negative control groups was successful withhigh scores shown by the vehicle control and the untreated control. Theeffect of treatment with strain 675 was striking and the mice treatedwith strain 675 exhibited notably reduced disease incidence and diseaseseverity. Indeed, the reduction in disease incidence and diseaseseverity was comparable to the positive control group. These dataindicate the strain 675 may be useful for treating or preventingmultiple sclerosis.

Example 7 Stability Testing

A composition described herein containing at least one bacterial straindescribed herein is stored in a sealed container at 25° C. or 4° C. andthe container is placed in an atmosphere having 30%, 40%, 50%, 60%, 70%,75%, 80%, 90% or 95% relative humidity. After 1 month, 2 months, 3months, 6 months, 1 year, 1.5 years, 2 years, 2.5 years or 3 years, atleast 50%, 60%, 70%, 80% or 90% of the bacterial strain shall remain asmeasured in colony forming units determined by standard protocols.

Sequences(Bacteroides coprocola gene for 16S rRNA, partial sequence, strain: M11 - AB200223) SEQ ID NO: 1 1agagtttgat cctggctcag gatgaacgct agctacaggc ttaacacatg caagtcgagg 61ggcagcatga acttagcttg ctaagtttga tggcgaccgg cgcacgggtg agtaacacgt 121atccaacctt ccgtttactc agggatagcc tttcgaaaga aagattaata cctgatagta 181tggtgagatt gcatgatggc accattaaag atttattggt aaacgatggg gatgcgttcc 241attaggtagt aggcggggta acggcccacc tagcctgcga tggatagggg ttctgagagg 301aaggtccccc acattggaac tgagacacgg tccaaactcc tacgggaggc agcagtgagg 361aatattggtc aatgggcgag agcctgaacc agccaagtag cgtgaaggat gaaggtccta 421cggattgtaa acttctttta tacgggaata aagtttccta cgtgtaggat tttgtatgta 481ccgtatgaat aagcatcggc taactccgtg ccagcagccg cggtaatacg gaggatgcga 541gcgttatccg gatttattgg gtttaaaggg agcgcagacg ggagattaag tcagttgtga 601aagtttgcgg ctcaaccgta aaattgcagt tgatactggt ttccttgagt gcagttgagg 661caggcggaat tcgtggtgta gcggtgaaat gcttagatat cacgaagaac cccgattgcg 721aaggcagctt gctaaactgt aactgacgtt catgctcgaa agtgtgggta tcaaacagga 781ttagataccc tggtagtcca cacggtaaac gatggatact cgctgttggc gatatactgt 841cagcggccaa gcgaaagcat taagtatccc acctggggag tacgccggca acggtgaaac 901tcaaaggaat tgacgggggc ccgcacaagc ggaggaacat gtggtttaat tcgatgatac 961gcgaggaacc ttacccgggc ttaaattgca gacgaattac gaggaaactt gtaagccgca 1021aggcgtctgt gaaggtgctg catggttgtc gtcagctcgt gccgtgaggt gtcggcttaa 1081gtgccataac gagcgcaacc ctcgtggtca gttactaaca ggttaagctg agggctctgg 1141ccagactgcc atcgtaagat gtgaggaagg tggggatgac gtcaaatcag cacggccctt 1201acgtccgggg ctacacacgt gttacaatgg gaggtacaga aggccgctac ccggcaacgg 1261gatgccaatc cccaaaacct ctctcagttc ggactggagt ctgcaacccg actccacgaa 1321gctggattcg ctagtaatcg cgcatcagcc acggcgcggt gaatacgttc ccgggccttg 1381tacacaccgc ccgtcaagcc atgaaagccg ggggtacctg aagtgcgtaa ccgcaaggag 1441cgccctaggg taaaaccggt aattggggct aagtctaaca aggtaaccaa g(Bacteroides coprocola gene for 16S rRNA, partial sequence, strain: M16 - AB200224) SEQ ID NO: 2 1agagtttgat cctggctcag gatgaacgct agctacaggc ttaacacatg caagtcgagg 61ggcagcatga acttagcttg ctaagtttga tggcgaccgg cgcacgggtg agtaacacgt 121atccaacctt ccgtttactc agggatagcc tttcgaaaga aagattaata cctgatagta 181tggtgagatt gcatgatggc accattaaag atttattggt aaacgatggg gatgcgttcc 241attaggtagt aggcggggta acggcccacc tagcctgcga tggatagggg ttctgagagg 301aaggtccccc acattggaac tgagacacgg tccaaactcc tacgggaggc agcagtgagg 361aatattggtc aatgggcgag agcctgaacc agccaagtag cgtgaaggat gaaggtccta 421cggattgtaa acttctttta tacgggaata aagtttccta cgtgtaggat tttgtatgta 481ccgtatgaat aagcatcggc taactccgtg ccagcagccg cggtaatacg gaggatgcga 541gcgttatccg gatttattgg gtttaaaggg agcgcagacg ggagattaag tcagttgtga 601aagtttgcgg ctcaaccgta aaattgcagt tgatactggt ttccttgagt gcagttgagg 661caggcggaat tcgtggtgta gcggtgaaat gcttagatat cacgaagaac cccgattgcg 721aaggcagctt gctaaactgt aactgacgtt catgctcgaa agtgtgggta tcaaacagga 781ttagataccc tggtagtcca cacggtaaac gatggatact cgctgttggc gatatactgt 841cagcggccaa gcgaaagcat taagtatccc acctggggag tacgccggca acggtgaaac 901tcaaaggaat tgacgggggc ccgcacaagc ggaggaacat gtggtttaat tcgatgatac 961gcgaggaacc ttacccgggc ttaaattgca gacgaattac gaggaaactt gtaagccgca 1021aggcgtctgt gaaggtgctg catggttgtc gtcagctcgt gccgtgaggt gtcggcttaa 1081gtgccataac gagcgcaacc ctcgtggtca gttactaaca ggttaagctg agggctctgg 1141ccagactgcc atcgtaagat gtgaggaagg tggggatgac gtcaaatcag cacggccctt 1201acgtccgggg ctacacacgt gttacaatgg gaggtacaga aggccgctac ccggcaacgg 1261gatgccaatc cccaaaacct ctctcagttc ggactggagt ctgcaacccg actccacgaa 1321gctggattcg ctagtaatcg cgcatcagcc acggcgcggt gaatacgttc ccgggccttg 1381tacacaccgc ccgtcaagcc atgaaagccg ggggtacctg aagtgcgtaa ccgcaaggag 1441cgccctaggg taaaaccggt aattggggct aagtctaaca aggtaaccaa(Bacteroides coprocola gene for 16S rRNA, partial sequence, strain: M158 - AB200225) SEQ ID NO: 3 1agagtttgat cctggctcag gatgaacgct agctacaggc ttaacacatg caagtcgagg 61ggcagcatga acttagcttg ctaagtttga tggcgaccgg cgcacgggtg agtaacacgt 121atccaacctt ccgtttactc agggatagcc tttcgaaaga aagattaata cctgatagta 181tggtgagatt gcatgatagc accattaaag atttattggt aaacgatggg gatgcgttcc 241attaggtagt aggcggggta acggcccacc tagcctncga tggatagggg ttctgagagg 301aaggtccccc acattggaac tgagacacgg tccaaactcc tacgggaggc agcagtgagg 361aatattggtc aatgggcgag agcctgaacc agccaagtag cgtgaaggat gaaggtccta 421cggattgtaa acttctttta tacgggaata aagtatccta cgtgtaggat tttgtatgta 481ccgtatgaat aagcatcggc taactccgtg ccagcagccg cggtaatacg gaggatgcga 541gcgttatccg gatttattgg gtttaaaggg agcgcagacg ggagattaag tcagttgtga 601aagtttgcgg ctcaaccgta aaattgcagt tgatactggt ttccttgagt gcagttgagg 661caggcggaat tcgtggtgta gcggtgaaat gcttagatat cacgaagaac cccgattgcg 721aaggcagctt gctaaactgt aactgacgtt catgctcgaa agtgtgggta tcaaacagga 781ttagataccc tggtagtcca cacggtaaac gatggatact cgctgttggc gatatactgt 841cagcggccaa gcgaaagcat taagtatccc acctggggag tacgccggca acggtgaaac 901tcaaaggaat tgacgggggc ccgcacaagc ggaggaacat gtggtttaat tcgatgatac 961gcgaggaacc ttacccgggc ttaaattgca gacgaattac gaggaaactt gtaagccgca 1021aggcgtctgt gaaggtgctg catggttgtc gtcagctcgt gccgtgaggt gtcggcttaa 1081gtgccataac gagcgcaacc ctcgtggtca gttactaaca ggttaagctg aggactctgg 1141ccagactgcc atcgtaagat gtgaggaagg tggggatgac gtcaaatcag cacggccctt 1201acgtccgggg ctacacacgt gttacaatgg gaggtacaga aggcagctac ccggcgacgg 1261gatgccaatc cccaaaacct ctctcagttc ggactggagt ctgcaacccg actccacgaa 1321gctggattcg ctagtaatcg cgcatcagcc acggcgcggt gaatacgttc ccgggccttg 1381tacacaccgc ccgtcaagcc atgaaagccg ggggtacctg aagtgcgtaa ccgcaaggag 1441cgccctaggg taaaaccggt aattggggct aagtcgtaac aaggtaacca a(consensus 16S rRNA sequence for Bacteroides coprocola strain 675)SEQ ID NO: 4GTCTGGCTCAKGATGAACGCTAGCTACAGGCTTAACACATGCAAGTCGAGGGGCAGCATGAACTTAGCTTGCTAAGTTTGATGGCGACCGGCGCACGGGTGAGTAACACGTATCCAACCTCCCGCTTACTCAGGAATAGCCTTTCGAAAGAAAGATTAATGCCTGATGGTATCTTAAGCACACATGTAATTAAGATTAAAGATTTATCGGTAAGCGATGGGGATGCGTTCCATTAGGTAGTAGGCGGGGTAACGGCCCACCTAGCCGACGATGGATAGGGGTTCTGAGAGGAAGGTCCCCCACATTGGAACTGAGACACGGTCCAAACTCCTACGGGAGGCAGCAGTGAGGAATATTGGTCAATGGGCGCGAGCCTGAACCAGCCAAGTAGCGTGAAGGATGAAGGTCCTATGGATTGTAAACTTCTTTTATACGGGAATAAAGTGGTCCACGTGTGGGCCTTTGCATGTACCGTATGAATAAGCATCGGCTAACTCCGTGCCAGCAGCCGCGGTAATACGGAGGATGCGAGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGCAGACGGGGGATTAAGTCAGTTGTGAAAGTTTGCGGCTCAACCGTAAAATTGCAGTTGATACTGGTTCCCTTGAGTGCAGTTGAGGCAGGCGGAATTCGTGGTGTAGCGGTGAAATGCATAGATATCACGAAGAACCCCGATTGCGAAGGCAGCCTGCTAAGCTGTAACTGACGTTGAGGCTCGAAAGTGTGGGTATCAAACAGGATTAGATACCCTGGTAGTCCACACGGTAAACGATGGATACTCGCTGTTGGCGATATACTGTCAGCGGCCAAGCGAAAGCATTAAGTATCCCACCTGGGGAGTACGCCGGCAACGGTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGAGGAACATGTGGTTTAATTCGATGATACGCGAGGAACCTTACCCGGGCTTAAATTGCAGACGAATTACTTGGAAACAGGTAAGCCGCAAGGCGTCTGTGAAGGTGCTGCATGGTTGTCGTCAGCTCGTGCCGTGAGGTGTCGGCTTAAGTGCCATAACGAGCGCAACCCTCGTGGCCAGTTACTAGCAGGTAACGCTGAGGACTCTGGCCAGACTGCCATCGTAAGATGCGAGGAAGGTGGGGATGACGTCAAATCAGCACGGCCCTTACGTCCGGGGCTACACACGTGTTACAATGGGAGGTACAGAAGGCAGCTACCCGGCGACGGGATGCCAATCTCCAAAGCCTCTCTCAGTTCGGACTGGAGTCTGCAACCCGACTCCACGAAGCTGGATTCGCTAGTAATCGCGCATCAGCCACGGCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCAAGCCATGAAAGCCGGGAGTACCTGAAGTGCGTAACCGCAAGGAGCGCCCTAGGGTAAAACCGGTAATTGGGGCTAAGTCNTACGGGG(Bacteroides thetaiotaomicron gene for 16S rRNA, partial sequence - M58763)SEQ ID NO: 5 1cttntacaat gaagagtttg atcctggctc aggatnaacg ctagctacag gcttaacaca 61tgcaagtcna ggggcagcat ttcagtttgc ttgcaaactg gagatggcga ccggcgcacg 121ggtgagtaac acgtatccaa cctgccgata actcggggat agcctttcga aagaaagatt 181aatacccnat ggtataatca gaccgcatng tcttrttatt aaagaatttc ggttatcgat 241ggggatgcgt tccattaggc agttggtgag gtaacggctc acnnaacctt cgatggatag 301gggttctgag aggaaggtcc cccacattgg aactgagaca cggtccaaac tcctacggga 361ggcagcagtg aggaatattg gtcaatgggc gcaggcctga accagccaag tagcgtgaag 421gatgactgcc ctatgggttg taaacttctt ttatatggga ataaagtttt ccacgtgtgg 481aattttgtat gtaccatatg aataaggatc ggctaactcc gtgccagcag ccncgntnat 541acggagnatc cgagcgttat ccggatttat tgggtttaaa gggagcgtag gtggacagtt 601aagtcagttg tgaaagtttg cggctcaacc gtaaaattgc agttgatact ggctgtcttg 661agtacagtag aggtgggcgg aattcgtggt gtagcggtga aatgcttaga tatcacgaag 721aactccgatt gcgaaggcag ctcactggac tgcaactgac actgatgctc gaaagtgtgg 781gtatcaaaca ggattagata ccctggtagt ccacacagta aacgatgaat actcgctgtt 841tgcgatatac agtaagcggc caagcgaaag cattaagtat tccacctggg gagtacgccg 901gcaacggtga aactcaaagg aattgacggg ggccngcaca agcggaggaa catgtggttt 961aattcgatga tacgcgagga accttacccg ggcttaaatt gcatttgaat atattggaaa 1021cagtatagcc gyaaggcaaa tgtgaaggtg ctgcatggtt gtcgtcagct cgtgccgtga 1081ggtgtcggct taagtgccat aacgagcgca acccttatct ttagttacta acaggtcatg 1141ctgaggactc tagagagact gccgtcgtaa gatgtgagga aggtggggat gacgtcaaat 1201cagcacngcc cntacgtccg gggctacaca cgtgttacaa tggggggtac agaaggcagc 1261tacctggtga caggatgcta atcccaaaag cctctctcag ttcggatcga agtctgcaac 1321ccgacttcgt gaagctggat tcgctagtaa tcgcgcatca gccatggcgc ggtgaatacg 1381ttcccgggcn ttgtacacac cgcccgtcaa gccatgaaag ccgggggtac ctgaagtacg 1441taaccgcaag gagcgtccta gggtaaaact ggtaattggg gc SEQ ID NO: 6(strain 675 chromosome sequence) - see electronic sequence listing.SEQ ID NO: 7(strain 675 plasmid sequence) - see electronic sequence listing.

REFERENCES

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The invention claimed is:
 1. A method of treating a conditioncharacterized by an elevated level of an IL-17 cytokine in a subject inneed thereof, comprising administering to said subject a pharmaceuticalcomposition that comprises a therapeutically effective amount of abacteria strain that comprises a 16S rRNA gene sequence with at least95% sequence identity to the polynucleotide sequence of SEQ ID NO:1, asdetermined by a Smith-Waterman homology search algorithm using an affinegap search with a gap open penalty of 12, a gap extension penalty of 2,and a Blocks Substitution Matrix (BLOSUM) of 62, wherein saidpharmaceutical composition is formulated for delivery to an intestine ofsaid subject, and wherein said administering is effective to treat saidcondition.
 2. The method of claim 1, wherein said condition is aninflammatory condition.
 3. The method of claim 2, wherein saidinflammatory condition is uveitis.
 4. The method of claim 3, whereinsaid uveitis is selected from the group consisting of Fuchsheterochromic iridocyclitis, HLA-B27 related uveitis, posterior uveitis,and uveitis syndrome.
 5. The method of claim 2, wherein saidinflammatory condition is arthritis.
 6. The method of claim 5, whereinsaid arthritis is selected from the group consisting of rheumatoidarthritis, osteoarthritis, psoriatic arthritis, spondyloarthritis,ankylosing spondylitis, and juvenile idiopathic arthritis.
 7. The methodof claim 2, wherein said inflammatory condition is a chronicinflammatory condition.
 8. The method of claim 2, wherein saidinflammatory condition is not responsive to treatment with a TNFαinhibitor.
 9. The method of claim 1, wherein said IL-17 cytokine isselected from the group consisting of: IL-17A, IL-17B, IL-17C, IL-17D,IL-17E, and IL-17F.
 10. The method of claim 1, wherein saidpharmaceutical composition further comprises a pharmaceuticallyacceptable excipient, diluent, or carrier.
 11. The method of claim 1,wherein said pharmaceutical composition comprises from about 1×10³ toabout 1×10¹¹ CFU/g of said bacteria strain, with respect to a totalweight of said pharmaceutical composition.
 12. The method of claim 1,wherein said pharmaceutical composition is encapsulated.
 13. The methodof claim 1, wherein said pharmaceutical composition is formulated as asuppository.
 14. The method of claim 1, further comprising administeringan additional therapeutic agent to said subject.
 15. The method of claim1, wherein said bacteria strain comprises a polynucleotide sequence of a16S rRNA gene that has at least 98% sequence identity to thepolynucleotide sequence of SEQ ID NO:1, as determined by aSmith-Waterman homology search algorithm using an affine gap search witha gap open penalty of 12, a gap extension penalty of 2, and a BlocksSubstitution Matrix (BLOSUM) of
 62. 16. The method of claim 1, whereinsaid bacteria strain comprises a polynucleotide sequence of a 16S rRNAgene that has at least 99% sequence identity to the polynucleotidesequence of SEQ ID NO:1, as determined by a Smith-Waterman homologysearch algorithm using an affine gap search with a gap open penalty of12, a gap extension penalty of 2, and a Blocks Substitution Matrix(BLOSUM) of
 62. 17. The method of claim 1, wherein said bacteria straincomprises the polynucleotide sequence of SEQ ID NO:
 1. 18. The method ofclaim 1, wherein said bacteria strain is lyophilized.
 19. The method ofclaim 1, wherein the condition is cancer.